Download A. Classical Adjuvants (Gel Suspensions, FDA Approved, used in

The World of Vaccine Adjuvants-The Hidden Secrets of Vaccine
The word ‘adjuvant’ is derived from the Latin word ‘adjuvare’ which means ‘to help’. Immunologic Adjuvants are added to vaccines to stimulate the
immune system's response to the target antigen, but do not in themselves confer immunity. Adjuvants accomplish this task by mimicking specific
sets of evolutionarily conserved molecules, so called PAMPs, which include liposomes, lipopolysaccharide (LPS), molecular cages for antigen,
components of bacterial cell walls (e.g., fl2000lins), and endocytosed nucleic acids such as double-stranded RNA (dsRNA), single-stranded DNA
(ssDNA), and unmethylated CpG dinucleotide-containing DNA (ODNs). Natural proteins such as ovalbumin or OVA-peptides and keyhole limpet
hemocyanins (KLH) are also being explored not only serve as carrier protein but also as adjuvants. Adjuvants are hidden secrets of vaccine.
ADI offers wide variety adjuvants, ELISA kits to monitor antigen specific antibody isotype responses in animals (mouse or rat, IgA, IgG1-4, IgM, IgE)
or humans. Most common adjuvants from various categories are also offered as convenient combo package containing 3-4 different adjuvants.
Separate ELISA kits are available to measure the antibodies to the adjuvant itself (KLH, Ovalbumin, Tetanus, Cholera, Diphtheria toxin, etc). We
also have ELISA kits to measure various vaccine components such as Tetanus, Diphtheria, Pertussis Toxin/Toxoid free or when adsorbed on Alum.
Some common types for adjuvants are:
A.
Alum or gel suspension as exemplified by Alumimum hydroxide (Alum/Alhydrogel)
B.
Biodegradable polymers or nanoparticle Adjuvants as exemplified by PLGA polymers or nanoparticles
C.
Emulsion/oil based adjuvants as exemplified by Freunds (complete and incomplete), Squalene, and Saponins
D.
Highly antigenic proteins toxins, peptides, synthetic chemical and DNA (animal, bacterial origins or synthetic)- as exemplified by LPS,
MPL, MPD, TDB
E.
Natural non-toxic proteins and bacteria toxins (animal, bacterial origins or synthetic) such as Albumins, KLH, Toxins and Toxoids
F.
Mammalian or bacterial DNA (single or double stranded) and Synthetic ODNs.
Not all vaccines contain Alum because an adjuvant may not have been needed, was not expected to increase the desired immune response, or was
going to cause an imbalance in the immune response. For example, inactivated Polio Virus (IPV/IPOL) vaccine, measles, mumps and rubella
vaccine (MMR/MMRII/MMRV), Varicella or chickenpox vaccine (Varivax/Proquad/MMRV), Meningococcal conjugate (MCV4/Menomune/Menactra)
vaccine, and influenza vaccines (Fluzone/Flulaval/Flumist/Fluvirin etc) do not contain aluminum salts.
A. Classical Adjuvants (Gel Suspensions, FDA Approved, used in approved vaccines)
Alum or gel suspension as exemplified by Aluminum hydroxide (Alum/Alhydrogel) or Aluminum phosphate (Adjuphos) or Calcium phosphate.
These gels non-specifically adsorb vaccine components such as proteins. Alum composition provides differences in net charge. Some proteins
may adsorb better on Alum, Adjuphos or Calcium phosphate. Alum-based adjuvants remain the only category of adjuvants that have been used in
humans and animals for more than 50-years. Many common vaccines such as DPT, hepatitis, rabies and anthrax vaccines use Alum and commonly
referred as adsorbed vaccine. The role of Alum is to provide stability to the protein and allow slow release and thereby enhance antibody
response. Vaccines formulated in Alum forms a depot at the injection site, enabling enhanced antigen availability, activation of antigen presenting
cells (APCs) and uptake by immune cells. Mostly Th2 (IgG1) response. Alum is non-toxic but only invokes a week response and antibody response
is typically Th2 or IgG1. The effectiveness of each salt as an adjuvant depends on the characteristics of the specific vaccine and how the
manufacturer prepares the vaccine. To work as an adjuvant, the antigen must be adsorbed to the Alum to keep the antigen at the site of injection.
Adjuvant
Aluminum Hydroxide,
Gels, Al (OH)3
Response
Th2 response
Cat # (Size)
AV-1010-100 (100 ml)
AV-1010-250 (250 ml)
Notes
Aluminum hydroxide Al(OH)3 or Alum) is wet gel suspension. low
endotoxin, FDA Approved, used in >80% approved vaccines; high
capacity non-specific adsorption of protein; good antibody response
independent of TLR signaling.
Issues: Mostly Th2 (IgG1) response; poor CD8 T-cell induction;
may cause mild local reaction and occasional granulomas.
Aluminum Phosphate
Gels; Al (PO4)3
Th2 response
AV-1015-100 (100 ml)
AV-1015-250 (250 ml)
ADJU-PHOS Al(PO4)3 is a wet gel suspension; low endotoxin,
FDA Approved, used in most human vaccines; high capacity nonspecific proteins adsorbed, Mostly Th2 (IgG1) response. Due to its
net –ve charge, it is used for positively charged proteins and
vaccines
Issues: Mostly Th2 (IgG1) response; poor CD8 T-cell induction;
may cause mild local reaction and occasional granulomas.
Calcium Phosphate
Th2 response
AV-1020-100 (100 ml)
AV-1020-250 (250 ml)
Calcium Phosphate is a wet gel suspension; low endotoxin, high
capacity non-specific proteins adsorbed, Mostly Th2 (IgG1)
response. Due to its net –ve charge, it is used for positively charged
proteins and vaccines; used in diphtheria, tetanus, pertussis and
poliomyelitis; some vaccine (Diphtheria) respond better than Alum;
An alternative to Alum.
Alum Gel
Combo, Trial Pak-1
Th2 response
AV-2000-PK-1
(10 ml x3)
Alum Gel Combo Trial Pak-1 Contains 10 ml each of Alhydrogel
(#AV-1010-10); Adjuphos (#AV-1015-10); and Calcium Phosphate
gel (#AV-1020-10) for testing various colloidal gels.
Alpha Diagnostic Intl Inc., USA; (800) 786-5777; [email protected]
;
www.4adi.com
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B
Biodegradable polymers or nanoparticle Adjuvants
These adjuvants as exemplified by PLGA polymers or nanoparticles. This is a new class of synthetic polymers varying in composition and
composition. The polymers assume the shape of nanosphere or nanoparticles. Vaccine components are encapsulated within the nanoparticles.
PLGA [poly(lactic-co-glycolic) acid] is the most attractive polymer for controlled or slow drug/vaccine delivery. It is a biodegradable FDA-approved
polymer of glycolic and lactic acid. PLGA 50:50 refers the ratio of 50% lactic and 50% glycolic acid. PLGA (75:25; 50:50, and 25:50%) are also
available. PLGA with end capped (esters) have longer half-lives ~6 months) than non-modified (~2 months). PLGA invokes Th1/2 response better
than Alum. Lupro anti-cancer drug is FDA approved that contains PLGA. PLGA emulsion produces W/O/W (water-in-oil-in-water) emulsions
Adjuvant
PLGA poly(lactic-coglycolic) acid); synthetic
Biodegradable
/Nanoparticles
Response
Th1/2
response
PLGA
Th1/2
response
Th1/2
response
Th1/2
response
Th1/2
response
PLGA
PLGA
PLGA
Combo Trial Pak-1
PLGA
PLGA-Glucose;
Poly(lactide-co-glycolide)
star, glucose core, hydroxyl
terminated
PLGA-PEG; Poly(lactideco-glycolide)-blockpoly(ethylene glycol)block-poly(lactide-coglycolide)
PLGA-Ester; Poly(ethylene
glycol) methyl etherblock-poly(lactide-coglycolide)
Cat # (Size)
Notes
PLGA is the most attractive polymer for controlled or slow drug/vaccine delivery. It is a
biodegradable FDA-approved polymer of glycolic and lactic acid. PLGA 50:50 refers the ratio of
50% lactic and 50% glycolic acid. PLGA (75:25; 50:50, and 25:50%) are available. PLGA with
end capped (esters) have longer half-lives ~6 months) than non-modified (~2 months). Invokes
Th1/2 response better than Alum. Lupro anti-cancer drug is FDA approved that contains PLGA.
PLGA emulsion produce W/O/W (water-in-oil-in-water) emulsions. Ongoing trials for Tetanus
and other vaccines: Issues: Difficult to prepare GMP formulations.
AV-2010-1 (1 g)
Contains 50% lactic and 50% glycolic acid, MW 30-60 Kda
AV-2020-1 (1 g)
Contains 65% lactic and 35% glycolic acid, MW 40-75 Kda
AV-2030-1 (1 g)
Contains 75% lactic and 25% glycolic acid, MW 65-105 Kda
AV-2000-PK-1
(100 mg x3)
PLGA-Trial Pak-1 Contains 100 mg each of the 3 common PLGA
(#AV-2010-10; AV-2020-1, and AV-2030-1). Designed to provide
various formulations of PLGA for testing adjuvantcy with a given
antigen
Contains 75% lactic and 25% glycolic acid; Ester capped, , MW 75115 Kda
contains LA:GA (50:50), hydroxyl terminated, degradation time 3-6
weeks; mw=~50 kda
Th1/2
response
Th1/2
response
AV-2040-1 (1 g)
Th1/2
response
AV-2060-1 (1 g)
typical PEG PDI<1.1; overall PDI<1.2 (THF, PEO); lactide:glycolide
25:75; PEG average Mn 1,000; PLGA average Mn 2000; average Mn
(1,000-1,000-1,000); degradation timeframe 1-2 weeks
Th1/2
response
AV-2070-1(1 g)
typical PEG PDI<1.1; overall PDI<1.8 (THF, PEO); lactide:glycolide
50:50; mol wt PEG average Mn 2,000; PLGA average Mn 15,000;
average Mn 17,000 (total); degradation time, 1-4 weeks
AV-2050-1 (1 g)
B. Classical Adjuvants (Oils/Emulsion type, used in most animal vaccines)
This class of adjuvants is exemplified by Freunds (complete and incomplete), Squalene, and Saponins. The adjuvants generally produce thick
water-in-oil emulsions. The primary objective is to delay the release or degradation of vaccines in the animals or humans. Generates an intense
inflammatory reaction at the site of antigen deposition. The adjuvant type and presence or other additives such as toxins or bacterial components
may further augment the antibody response. Depending upon the adjuvant, antibody response is Th1, Th2 or both. These adjuvants are primarily
used in animal vaccines or experimental vaccines
Adjuvant
(CFA) Complete
Freund’s Adjuvant
Response
Th1/2
response.
Cat # (Size)
AV-3010-10 (10 ml)
AV-3010-100 (10x10ml)
IFA (Incomplete
Freund’s adjuvant)
Th2 response
AV-3015-10 (10 ml)
AV-3015-100 (10x10ml)
Squalene
Th2 response
AV-3020-10 (10 ml)
Classical Adjuvant
Combo Pak-1
AV-3000-PK-1 (10 ml x3)
Notes
Contains heat-killed M. tuberculosis, Mannide Monoleate and
Paraffin oil. It is water-in-oil emulsion. Produced high titer
antibodies (IgG1-4/IgM and other isotypes). Primarily used as
experimental adjuvant and only as the first vaccine dose. Issues:
strong reaction at the injection site. Not recommended for vaccines.
Used to assess and compare other adjuvants.
Same as CFA except M. tuberculosis. It is water-in-oil emulsion.
Generates an intense inflammatory reaction at the site of antigen
deposition. Used in conjunction with CFA; all injections can be
given in IFA after the initial dose with CFA. Issues: May cause
granulomas.
Squalene-based oil-in-water nano-emulsions promote a significant
increase in antibody titers with reportedly more balanced Th1/Th2
responses than those obtained with alum. Increased performance
for flu, HBV, HSV, HIV, and CMV experimental vaccines. Issues:
mild local reaction and mild reactogenicity when used intramuscular.
Contains 3 most common classical oil-based adjuvant emulsions;
10 ml each of Complete Freund’s Adjuvant/CFA (#AV-3010-10);
Incomplete Freund’s adjuvant/IFA (AV-3015-10), and Squalene
(#AV-3020-10) to test adjuvantcy with a given antigen
Alpha Diagnostic Intl Inc., USA; (800) 786-5777; [email protected]
;
www.4adi.com
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C. Classical Adjuvants (Oils/Emulsion type, used in most animal vaccines)
Adjuvant
Saponins (Quillaja bark)
Saponins
Combo Pak-1
Response
Th1/2
response
Cat # (Size)
AV-4010-1 (1 mg)
AV-4010-10 (10 mg)
Th1/2
response
AV-4020-100 (100 mg)
AV-4020-1000 (1 g)
Th1/2
response
AV-4030-10 (10 g)
Th1/2
response
AV-4000-PK-1 (3 vials)
Peanut oil
AV-5010-50 (50 ml)
Mineral oil
AV-5020-50 (50 ml)
Mannide monooleate
AV-5030-5 (5 g)
Oil Adjuvant
Combo Pak-1
AV-5000-PK-1 (3 vials)
HS15 Kolliphore
AV-6010-100 (100g)
Notes
QS21 (Quil-A) Vaccine adjuvant, (EU GMP grade) >95%; Highly
potent adjuvant from Quillaja saponaria from the bark of Molina tree;
more balanced Th1/Th2 responses than those obtained with alum.
Used many veterinary and experimental vaccines.
Vet-SAP Vaccine adjuvant, (EU GMP grade) >95%; Highly potent
adjuvant from Quillaja saponaria from the bark of Molina tree; more
balanced Th1/Th2 responses than those obtained with alum. Used
many veterinary and experimental vaccines.
Saponin (Quillaja bark) pure, (sapogenin 25-35%); Nonionic
sufactant that forms oil-in-water emulsions and acts as protective
colloid
Saponin Combo Pak-1 contains 3 vials of most common
Saponins from (Quillaja bark); 1 mg of QS21 (#AV-4010-1); 100 mg
of Vet-SAP (#AV-4020-100) and 100 mg Saponin (#AV-4030-1);
These 3 Saponins differ in the purity and cost. Designed to test the
Adjuvantcy of Saponins with a given antigen.
Peanut oil, Delivery vehicle for fat-soluble compounds; Non-polar
solvent, largely consisiting of long-chain glycerides of fatty acids,
used for the delivery of lipophilic compounds.
Mineral oil functions by forming a depot at the injection site slowly
releasing antigen. Suitable for testing antigenicity of experimental
vaccines
Mannide monooleate; An emulsifying agent utilized in preparing
Freund′s incomplete adjuvant and to prepare oil-emulsion of
proteins; for animal studies
Oil Adjuvant Oil Pak-1 Contains 3 vials of: Peanut Oil (10 ml, #AV5010-10); Mineral oil (10 ml #AV-5020-10) and Mannide
monooleate (1 g #AV-5030-1)
Non-ionic solubilizer for manufacturing aqueous parenteral
preparations with lipophilic proteins and vaccines
D. Adjuvants or Additives (Used alone or with other adjuvants) Bacterial origins
These adjuvants also act as an irritant which causes the body to recruit and amplify its immune response. A tetanus, diphtheria, and pertussis
vaccine, for example, contains minute quantities of toxins/toxoids produced by each of the target bacteria. The body's immune system develops an
antitoxin to the bacteria's toxins as well as vaccine components. The vaccines that are poor antigen are generally formulated with the toxins. Many
Hib, pneumococcal vaccines are formulated in tetanus toxoid or diphtheria toxoid or other toxins.
Adjuvant
Recombinant Fl2000lin
FliC
(LPS)
Lipopolysaccharides
from Escherichia
coli 0111:B4
(LPS)
Lipopolysaccharides
from Salmonella enterica
serotype typhimurium
Lipopolysaccharides
from Escherichia
coli 0111:B4 ultra pure
Response
TLR5 Agonist
Th1/Th2
TLR4 agonist
Cat # (Size)
AV-7010-50 (50 ug)
AV-7015-1 (1 mg)
AV-7015-10 (1 mgx10)
TLR4 agonist
AV-7020-1 (1 mg)
AV-7020-10 (1 mgx10)
TLR4 agonist
AV-7016-1 (1 mg)
Lipopolysaccharides
from Escherichia
coli 0111:B4 ultra pure
Monophosphoryl lipid A
(MPLA)-SM
MPLA from S. enterica
Minnesota
MPLA Synthetic
TLR4 agonist
AV-7016-5 (5 mg)
TLR4 agonist
Th1 response
AV- 7025-1 (1 mg)
AV- 7025-5 (5 mg)
MDP, muramyldipetide
(Ac-muramyl-Ala-D-Gluamide) Synthetic
Diphosphoryl Lipid A (E.
coli K12 D31m4)
TLR2
AV-7035-1 (1 mg)
AV-7035-5 (5 mg)
TLR4 agonist
AV-7040-1 (1 mg)
AV-7040-5 (5 mg)
TLR4 agonist
Th1 response
AV-7030-1
AV-7030-1
Notes
Bacterial fl2000lin, a TLR5 ligand, is a potent T-cell antigen and has
potential as a vaccine adjuvant in flu vaccines.
LPS is a major constituent of the cell wall of most gram negative
bacteria. It is a highly immunogenic antigen with the ability to
enhance immune responses to soluble antigens in most animals.
Issues: endotoxins and reaction to E. coli derived LPS.
LPS is a major constituent of the cell wall of most gram negative
bacteria. It is a highly immunogenic antigen with the ability to
enhance immune responses to soluble antigens in most animals
LPS is a major constituent of the cell wall of most gram negative
bacteria. It is a highly immunogenic antigen with the ability to
enhance immune responses to soluble antigens in most animals.
Issues: endotoxins and reaction to E. coli derived LPS.
LPS is a major constituent of the cell wall of most gram negative
bacteria. It is a highly immunogenic antigen with the ability to
enhance immune responses to soluble antigens in most animals..
Lipid A from Salmonella minnesota R595 lipopolysaccharide (LPS
or endotoxin). Less toxic than LPS. MPLA-SM is a potent activator
of TLR4; Issues: Endotoxin related issues and immune reaction to
LPS.
Synthetic lipid A from E-coli serotype R515 .Potent activator of
TLR4; No LPS or bacterial endotoxins; Detoxified version of MPLA
(from S. enterica). Component of Melanoma vaccine. Issues:
autoimmune and neuro-inflammatory disorders.
Synthetic peptide; a neurotrophic and immunomodulatory factor
related to bacterial peptidoglycans, used as an immune modulator;
Enhances antigenicity of weak antigens, increasing antibody titer.
Lipid A, a disaccharide with fatty acid side chains, recognized by
TLR4 receptors, is the component responsible for the endotoxic
activity of LPS
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Peptidoglycan from S.
aureus
works like MDP
LPS/MPLA/MDP
Combo Pak-1
TLR2
TLR2/TLR4
AV-7045-5 (5 mg)
AV-7045-25 (25 mg)
AV-7000-PK-1 (3 vials)
Synthetic peptide; a neurotrophic and immunomodulatory factor
related to bacterial peptidoglycans, used as an immune modulator;
Enhances antigenicity of weak antigens, increasing antibody titer.
LPS/MPLA/MDP Adjuvants Combo Pak1, contains 3x1 mg of LPS #
AV-7015-1), 1 mg of MPLA-SM (#AV- 7025-1) and 1 mg of MDP
(#AV-7035-1);
Adjuvants or Additives (Used alone or with other Adjuvants) Synthetic
These adjuvants also act as an irritant which causes the body to recruit and amplify its immune response. A tetanus, diphtheria, and pertussis
vaccine, for example, contains minute quantities of toxins/toxoids produced by each of the target bacteria. The body's immune system develops an
antitoxin to the bacteria's toxins as well as vaccine components. The vaccines that are poor antigen are generally formulated with the toxins. Many
Hib, pneumococcal vaccines are formulated in tetanus toxoid or diphtheria toxoid or other toxins.
Adjuvant
Gardiquimod/
Imidazoquinoline
Response
TLR7 agonist
Th1 response
Cat # (Size)
AV-8010-5 (5 mg)
AV-8010-25 (25 mg)
Notes
Induces the activation of NF-κB in HEK 293 cells expressing TLR7
Imiquimod/
Imidazoquinoline
TLR7 agonist
Th1 response
AV-8015-5 (5 mg)
AV-8015-25 (25 mg)
Induces production of pro-inflammatory cytokines, such as IFN—
gamma and IL-12; activation of both innate and acquired immunity.
R848/ Resiquimod
TLR7/8 agonist
Th1 response
AV-8020-1 (1 mg)
AV-8020-5 (5 mg
TLR7/8
AV-8000-PK-1
Synthetic Adjuvant
Combo Pak-1
potent synthetic agonist of TLR7/TLR8; antiviral and antitumor,
synergistic effect with poly(dT)
Synthetic Adjuvant Combo Pak-1 contains 3 vials: 5 mg of
Gardiquimod #AV-8010-5; 5 mg of Imiquimod (#AV-8015-5) and 1
mg of R848 (#AV-8020-1).
Potent synthetic agonist of TLR7. Induces immune cell activation
and increases cytokine production. Displays antitumor and antiviral
activity in various animal models.
Non-toxic synthetic analogue of the mycobacterial cell wall
component trehalose 6,6’ dimycolate (TDM, also known as cord
factor). most studied immunostimulatory component of
Mycobacterium tuberculosis
Toll-like receptor 2/6 (TLR2/6) agonist. Induces TNF-α production in
human mononuclear cells. Also induces proliferation and activation
of mouse splenic B cells
RWJ 21757
Synthetic
TLR7 agonist
Th1 response
AV-9010-1 (1 mg)
AV-9010-10 (5 mg
TDB (Trehalose-6,6dibehenate) Synthetic
Th1 response
Mincle agonist
AV-9015-2 (2 mg)
AV-9015-5 (5 mg)
Pam2CSK4
TLR2/6 agonist
Th1 response
AV-9020-1 (1 mg)
AV-9020-B (Biotin)
AV-9020-F (FITC)
Pam3CSK4
TLR1/2 agonist
Th1 response
AV-9025-1 (1 mg)
AV-9025-B (Biotin)
AV-9025-B (FITC)
Synthetic triacylated lipoprotein. Induces Reported to increase
antibody responses to flu antigens. TNF-α and IL-6 in macrophage
Poly(I:C) (PolyinosinicPolycytidylic) Synthetic
TLR3 agonist
Th1 response
AV-9030-10 (10 mg)
AV-9030-50 (50 mg)
Synthetic Adjuvant
Combo Pak-2
TLR1/2/3/6
agonists
Th1 response
AV-9000-PK-2 (3 vials)
Poly(I:C); Double-stranded homopolymer; TLR3 agonist Polyinosine-polycytidylic acid. Th1 response; potent adjuvant to
enhance the specific anti-tumor immune responses against a
peptide-based vaccine.
Synthetic Adjuvant Combo Pak-2 contains: 1 vial of Pam2csk4
(#AV-9020-1 (1 mg), 1 vials of Pam3csk4 (#AV-9025-1 (1 mg), and
1 vial of Poly(I:C) (#AV-9030-10 (10 mg)
Adjuvants or Additives (Used alone or with other Adjuvants): Natural Proteins and Toxoids
Many small molecules such as peptides, haptens, drugs, hormones or vaccine components (polysaccharides) are not antigenic or poorly antigenic.
These antigens must be conjugated to natural, nontoxic proteins carrier proteins such as Albumins (animal, human, avian origins), hemocyanins or
powerful bacterial endotoxins/toxoids (Diphtheria, Tetanus, Pertussis etc). Some known ovalbumin (OVA) peptides also stimulate immune
responses.
Adjuvant
KLH
Keyhole limpet
Hemocyanins
Ovalbumin Protein
Response
Th1/2
Cat # (Size)
AV-9305-10 (10 mg)
AV-9305-100 (100 mg)
Th1/2
AV-9310-1000 (1g)
Ovalbumin Peptide
OVA 257-264 class I
(MHC) molecule,
Th1/2
AV-9315-1 (1 mg)
AV-9315-10 (10 mg)
Notes
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations .
Used as model peptide to study antigenicity, adjuvants and vaccine
formulations
Ovalbumin Peptide
OVA 323-339 MHC
class II peptide-
Th1/2
AV-9320-1 (1 mg)
AV-9320-10 (10 mg)
Used as model peptide to study antigenicity, adjuvants and vaccine
formulations
Alpha Diagnostic Intl Inc., USA; (800) 786-5777; [email protected]
;
www.4adi.com
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Adjuvants or Additives (Used alone or with other Adjuvants): Natural Proteins and Toxoids
Many small molecules such as peptides, haptens, drugs, hormones or vaccine components (polysaccharides) are not antigenic or poorly antigenic.
These antigens must be conjugated to natural, nontoxic proteins carrier proteins such as Albumins (animal, human, avian origins), hemocyanins or
powerful bacterial endotoxins/toxoids (Diphtheria, Tetanus, Pertussis etc). Some known ovalbumin (OVA) peptides also stimulate immune
responses.
Adjuvant
DNP-KLH Conjugate
Response
Th1/2
Cat # (Size)
AV-9325-10 (10 mg)
Notes
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
DNP-BSA Conjugate
DNP-HSA Conjugate
Th1/2
AV-9330-BSA (10 mg)
AV-9330-HSA (10 mg)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
DNP-Ovalbumin
Conjugate
Th1/2
AV-9335-10 (10 mg)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
DNP-Lipopolysaccharide
(LPS) Conjugate
Th1/2
AV-9340-1 (1 mg)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Tetanus Toxoid
C. tetanii
Th1/2
AV-9105-25 (25 ug)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Rec. Tetanus Light
Chain, (Non-toxic)
Th1/2
AV-9110-10 (10 ug)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Tetanus Light Chain, Cterminal fragment
(Non-toxic)
Th1/2
AV-9115-10 (10 ug)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Diphtheria Toxoid
Th1/2
AV-9120-500 (500 ug)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Diphtheria Toxin
CRM197 (non-toxic)
Th1/2
AV-9125-100 (100ug)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Pertussis toxin
B. Pertussis
Th1/2
AV-9130-50 (50 ug)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Cholera toxin
Vibrio cholera
Th1/2
AV-9135-500 (500 ug)
AV-9135-1000 (1 mg)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Recombinant Anthrax
toxin PA83 (non-toxic)
Th1/2
AV-9140-100 (100ug)
AV-9140-1000 (1000ug)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Pasteurella multicoda
toxin
Th1/2
AV-9145-10 (10 ug)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Exotoxin A from P.
aeruginosa
Th1/2
AV-9150-10 (100 ug)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Bacterial endotoxins
Combo Pak-1
Th1/2
AV-9200-PK-1 3 vials
1 vial of Tetanus Toxoid (#AV-9105-N-25 (25 ug); 1 vial of
Diphtheria Toxin CRm197 (#AV-9125-100; 100ug) and 1 vial of AV9135-500 (500 ug)
Generally used as carrier protein to increase antigenicity of poorly
antigenic peptide/proteins, haptens, and drugs. Also used model
antigen to study adjuvant or vaccine formulations
Alpha Diagnostic Intl Inc., USA; (800) 786-5777; [email protected]
;
www.4adi.com
Page 5
of 8
CpG ODN Nucleotide/DNA Adjuvants- CpG oligodeoxynucleotides (or CpG ODN)
These adjuvants are short single-stranded synthetic DNA molecules that contain an unmethylated CG (Cytosine–guanine) di nucleotide in a specific
base sequence (CpG motifs). CpG oligodeoxynucleotides (ODNs) represent a novel pharmacotherapeutic class with profound immunomodulatory
properties. These CpG motifs are not seen in eukaryotic DNA, in which CG dinucleotides are suppressed and, when present, usually methylated,
due to this they are considered pathogen-associated molecular patterns (PAMPs). The CpG PAMP is recognized by the pattern recognition
receptor (PRR) Toll-Like Receptor 9 (TLR9), which is constitutively expressed only in B cells and plasmacytoid dendritic cells (pDCs) in humans and
other higher primates. Unmethylated CpG ODNs (18–24 bp in length) possess immunomodulatory properties similar to bacterial DNA and they act
as immuno stimulants. The immunostimulatory properties of CpGs include induction of a Th1-type response with prominent release of IFN-Á, IL-12,
and IL-18. . CpG ODNs are classified into 5 classes, based on their sequence, secondary structures, and effect on human peripheral blood
mononuclear cells (PBMCs). The five classes are Class A (Type D), Class B (Type K), Class C, Class P, and Class S. Class A stimulate the
production of large amounts of Type I interferons (IFNα) and induce the maturation of pDCs. They are also strong activators of NK cells through
indirect cytokine signaling. Class B ODN are strong stimulators of human B cell and monocyte maturation. Class C ODN combine features of both
types A and B. They induce strong IFN-α production from pDC and B cell stimulation. Inhibitory ODNs are new class of ODNs that suppress TLR
(Toll-like receptor) 9. Class I: G-stretch ODNs: TLR9-specific competitors, some iODNs may also affect TLR7 and TLR8 signaling. Class II: ODNs
with telomeric repeats: TLR-independent inhibitors of STAT signaling (cellular uptake via an “ODN receptor”?) Class III: Inhibitors of DNA uptake in a
sequence independent manner Class IV: Long phosphorothioate ODNs as direct competitors of TLR9 signaling in a sequence independent manner.
Items
Description
TLR
Type
Unlabeled
Cat #
Biotin Conj.
Cat #
Negative
Control*
ODN1585-1
ODN1585-5
ODN1668-1
ODN1668-5
ODN1826-1
ODN1826-5
ODN2006-1
ODN2006-5
ODN2007-1
ODN2007-5
ODN2216-1
ODN2216-5
ODN2336-1
ODN2336-5
ODN2395-1
ODN2395-5
ODNM362-1
ODNM362-5
ATODN1-1
ODN1585-B
4
ODN1585-1NC
ODN1585-5NC
ODN1668-1NC
ODN1668-5NC
ODN1826-1NC
ODN1826-5NC
ODN2006-1NC
ODN2006-5NC
ODN2007-1NC
ODN2007-5NC
ODN 2216-1NC
ODN 2216-5NC
ODN2336-1NC
ODN2336-5NC
ODN2395-1NC
ODN2395-5NC
ODNM362-1NC
ODNM362-5NC
4
ATODN2-1
4
4
ODN1585
ODN1585-Type A, Antigen grade
ODN 1668
ODN 1668- Type B, Antigen grade;
ODN 1826
ODN 1826- Type B, antigen grade
ODN 2006
ODN 2006-Type B-, antigen grade;
ODN 2007
ODN 2007-Type B antigen grade
ODN 2216
ODN 2216-Type A, antigen grade
ODN 2336
ODN 2336-Type A, antigen grade;
ODN 2395
ODN 2395-Type C, antigen grade.
ODN M362
ODN M362-Type C, antigen grade
AT-ODN-1
AT ODN-Non-CpG AT rich ODN, Antigen grade
AT-ODN-2
AT ODN-Non-CpG AT rich ODN. Antigen grade
TLR9
agonist
Murine
TLR9 agonist
Murine
TLR9 agonist
Human
TLR9 agonist
Bovine/Porcine
TLR9 agonist
Human TLR9
agonist
Human TLR9
agonist
Human TLR9
agonist
Human/murine
TLR9 agonist
Murine
TLR9 agonist
TLR9 agonist
AT-ODN-2
AT ODN-Non-CpG AT rich ODN. Antigen grade
TLR9 agonist
ATODN3-1
4
4
Neutral ODN
Control
ODN-NT-1
4
4
ODN006-1
4
4
ODN 2088
Control for inhibitory ODNs. Antigen grade, No
agonistic or Antagonist activity
ODNBW006 CpG ODN (type B sequence at 5’
and Type A at the 3' end), antigen grade;
ODN 2088- Class I, antigen grade
ODN 4084-Type B, antigen grade
4
ODN2088-1NC
ODN2088-5NC
4
ODN INH-1
ODN INH-1 -Class R (restricted) antigen grade
TLR9 Antagonist
ODN2088-1
ODN2088-5
ODN4084F-1
ODN4084F-5
ODNIHN1-1
4
ODN 4084-F
Murine TLR9
Antagonist
TLR9 Antagonist
4
4
ODN INH-47
ODN INH-47- Class R (restricted) antigen grade
TLR9 Antagonist
ODNINH47-1
4
4
ODN TTAGGG
ODN TTAGGG-Inhibitory ODN , Antigen grade
ODNTT-1
4
ODNTT-1NC
G-ODN
G-ODN Murine, Antigen grade
GODN-1
4
4
Core-iODN
Inhibitory ODN class I, antigen grade
CIODN-1
4
4
Super-iODN
SIODN-1
4
4
Bacterial DNA
Inhibitory ODN- class I/II hybrid (TLR7 and
TLR8), antigen grade
Bacterial Plasmid dsDNA, >95%, antigen grade
Human TLR9
antagonist
Murine TLR9
antagonist
TLR7/9
antagonist
TLR7/9
antagonist
Agonists
Mammalian
DNA
Mammalian
DNA
Purified Native dsDNA, Calf Thymus,
ELISA/Antigen grade
Purified Native ssDNA, Calf Thymus,
ELISA/Antigen grade
ODN BW006
Agonists
Agonists
ODN1668-B
ODN1826-B
ODN2006-B
ODN2007-B
ODN 2216-B
ODN2336-B
ODN2395-B
ODNM362-B
PDNA15-N100
PDNA15-N100
DNAS35-N-1
Notes: Negative control contains GpC nucleotides instead of CpG. 4. Contact ADI to place a special order for Biotin or FITC Conjugates.
Alpha Diagnostic Intl Inc., USA; (800) 786-5777; [email protected]
;
www.4adi.com
Page 6
of 8
CpG ODN Nucleotide/DNA Adjuvants-Combo Paks
Items
ODN Combo Pak-1
ODN Combo Pak-2
ODN Combo Pak-3
ODN Combo Pak-4
ODN Combo Pak-5
Description
Combo Pak
Cat#
TLR
Types
Mouse TLR9-agonist kit; contains 100 ug of ODN1585-1; ODN1826-1,
ODN2395-1, and negative controls ODN1585-1NC; ODN1826-1-1NC,
ODN2395-1NC
Human TLR9-agonist kit; contains 100 ug of ODN2006-1; ODN2216-1;
ODN2395-1; and negative controls ODN2006-1NC; ODN2216-1NC;
ODN2395-1NC
TLR9-antagonist kit; contains 100 ug of ODN2088-1; ODN4084-F;
ODNINH-1; ODNINH-18; ONDTT-1.
TLR9-A&C class agonist kit, Contains 100 ug of ODN1585-1; ODN22161; ODN2336-1;ODN 2395 and ODN M362; ODNDSL03
TLR9-B class agonist kit, Contains 100 ug of- ODN1668-1; ODN1826;
and ODN2006
ODN-PK-1
Mouse TLR9,
agonist
ODN-PK-2
Human TLR9,
agonist
ODN-PK-3
TLR9, antagonist
ODN-PK-4
TLR9-A&C class,
agonist
TLR9-B class,
agonist
ODN-PK-5
NOTES: ADI also provides packages of 1 mg or 5 mg (standard package) or custom bulk package. All ODNs are >95% HPLC pure, and <1 EU
Endotoxin, antigen grade. ADI can also custom design and make any ODN not listed in the catalog.
ELISA Kits for the study of Adjuvant effect on antibody response
The main function of adjuvants (conventional oil-based, special formulation or ODNs) is to enhance antibody response to a given antigen. The
antibody levels or titer in the host species (mouse, rat, rabbit, G. pig, Ferret, Hamster, monkey or human etc) is typically measure by ELISA. ADI
provides several species specific ELISA kits to measure antibody titer. These kits contain all necessary reagents, except user’s specific antigen, for
about 1000 tests. Please see details at:
http://4adi.com/commerce/catalog/spcategory.jsp?category_id=2473
ADI also has ELISA kit for IgG, IgM, IgA, IgE or individual IgG isotypes (IgG1, 2a, 2b, IgG3) in serum or plasma for mouse, rat, human etc.
http://4adi.com/commerce/catalog/spcategory.jsp?category_id=2446
ELISA Kits for the study of Adjuvant effect on antibody response using model antigens
Several model proteins have been used as model antigens (Bovine serum albumin/BSA or HSA, ovalbumin or OVA, Keyhole limpet
hemocyanins/KLH, thyroglobulin, DNP-KLH/Albumin or Tetanus Toxoid or TTX or Diphtheria Toxoids. Adjuvants (conventional or ODN-based) have
been used to study antibody response to a model antigen in a given species of host. ADI has specific antibody ELISA kits for these model antigens.
Please see available kits at:
http://4adi.com/commerce/catalog/spcategory.jsp?category_id=2719
http://4adi.com/commerce/catalog/spcategory.jsp?category_id=2449
Vaccine_Adjuvants_flr
140430A
Alpha Diagnostic Intl Inc., USA; (800) 786-5777; [email protected]
;
www.4adi.com
Page 7
of 8