ImmunEasy Mouse Adjuvant

For induction of high antibody titers in mice

Features and benefits

• Easy mixing and delivery in aqueous suspension
• Rapid production of high antibody titers
• Boosting performed after only 2 weeks
• Less antigen required than with conventional methods
• Non-toxic adjuvant with no adverse side effects

Principle

ImmunEasy Mouse Adjuvant is a new type of adjuvant specifically designed for inducing high antibody titers in mice. The adjuvant contains immunostimulatory CpG DNA — short oligonucleotides that contain unmethylated cytosine-guanine dinucleotides within a certain base context. The mammalian immune system has evolved to recognize these sequences, which are found naturally in bacterial DNA, as a sign of infection. Exposure to CpG DNA results in very rapid and strong immune activation, and, when applied with an antigen, CpG DNA produces high titers of antigen-specific antibodies.

The antibody titer induced by ImmunEasy Mouse Adjuvant is generally equal to or higher than that induced by Freund’s complete adjuvant (see figure "Induction of High Antibody Titers"). In addition, high antibody titers are induced using up to 10-fold less antigen compared to some other adjuvants. Furthermore, boosting can be performed after 2 weeks instead of 4 weeks, saving both time and animal-housing costs.

The immune response induced by CpG DNA is based on a natural reaction. Therefore, unlike many oil-based adjuvants, ImmunEasy Mouse Adjuvant does not induce symptoms of toxicity in immunized animals, even at doses much higher than those required for adjuvant activity (see figure "Less Muscle Damage Using ImmunEasy Mouse Adjuvant").

Induction of High Antibody Titers

Augmentation of anti-HBs titers by different adjuvants in BALB/c mice. Total IgG titers in plasma of BALB/c mice immunized by intramuscular injection of 1 µg hepatitis B surface antigen (HBsAg) alone (no adjuvant) or in combination with various adjuvants. Each bar represents the geometric group mean (n=5–10) for total IgG anti-HBs. (Data excerpted from Weeratna, R.D., McCluskie, M.J., Xu, Y., and Davis, H.L. [2000] CpG DNA induces stronger immune responses with less toxicity than other adjuvants. Vaccine 18, 1755–1762. Copyright 2000 with permission from Elsevier Science and the authors.)

Less Muscle Damage Using ImmunEasy Mouse Adjuvant

BALB/c mice were immunized with a single dose of 1 µg recombinant HBsAg alone (no adjuvant) or with different adjuvants. The muscles were removed and photographed seven days later. The degree of damage in the muscle was scored blind by nine evaluators who were shown several slides for each adjuvant in random order. Scores indicate 0: no, 1: minimal, 2: moderate, and 3: severe damage. The bars represent the mean score for muscle damage for each adjuvant. (Data excerpted from Weeratna, R.D., McCluskie, M.J., Xu, Y., and Davis, H.L. [2000] CpG DNA induces stronger immune responses with less toxicity than other adjuvants. Vaccine 18, 1755–1762. Copyright 2000 with permission from Elsevier Science and the authors.)

Procedure

ImmunEasy Mouse Adjuvant is provided as a ready-to-use aqueous suspension. The procedure is easy — adjuvant and antibody are mixed, incubated for 5–10 minutes at room temperature, and then injected into the mouse. Tedious mixing procedures such as heating, sonification, lyophilization, and homogenization, which are required for Freund’s adjuvant and other water-in-oil emulsions, are completely avoided, as are problems associated with improperly prepared adjuvant–antigen mixes. Boosting is performed 2 weeks after the primary immunization.

Applications

ImmunEasy Mouse Adjuvant can be used with most types of antigen, including peptides, soluble proteins, virus-like particles, whole cell preparations, and polysaccharide conjugates.

Since different CpG DNA sequences activate the immune systems of different animal species, ImmunEasy Mouse Adjuvant is specifically designed for immunization of mice. Use in other species, including other types of rodents, will not yield acceptable titers.

操作手册: ImmunEasy™ Mouse Adjuvant Handbook (PDF version, 69 KB)