Mouse Rapid Aqueous Adjuvant

Q1: How to choose MelonAntibody series immunization adjuvants?

Our water-soluble MelonAntibody series immunization adjuvants are available for various animals, including: water-based rapid immunization adjuvants for rabbits, mice, alpacas, goats, chickens, and pigs.

Q2: What are the characteristics of the MelonAntibody series immunization adjuvants?

MelonAntibody immunization adjuvant offers several advantages compared to conventional Freund's adjuvant, detailed as follows:

1. MelonAntibody requires only two immunization shots, whether for monoclonal or polyclonal antibody preparation, reducing the number of immunization shots compared to Freund's adjuvant.
2. By reducing the number of immunization shots and lowering the antigen dosage per shot, MelonAntibody significantly saves the total antigen dosage. Recommended antigen dosages are as follows: (1) For most protein antigens, 25-50μg per shot is suggested; (2) For highly immunogenic inactivated whole virus, whole bacteria, and virus-like particle antigens, 15-25μg per shot is recommended; (3) For small molecule antigens conjugated with carrier proteins, 50-80μg per shot is advised. Actual dosages should be primarily based on the company's previous data and experience.
3. MelonAntibody facilitates rapid antibody production with high titers and affinity. In a standard immunization protocol, whether for monoclonal or polyclonal antibody preparation, only two to three shots are required within three weeks, typically achieving ELISA titers (with a cutoff value of 0.1000) of 1:10000-1:10000000 by the fourth to fifth week.
4. MelonAntibody preserves the natural conformation of antigens, facilitating the screening and acquisition of monoclonal antibodies targeting conformational antigen epitopes, a critical feature lacking in Freund's adjuvant.
5. MelonAntibody is water-soluble, eliminating the need for the complex emulsification process required by Freund's adjuvant. Antigen and adjuvant only require simple mixing for animal immunization.
6. MelonAntibody utilizes the intramuscular immunization route, greatly simplifying the process compared to conventional mouse monoclonal antibody preparation, which may involve paw or splenic immunization.
7. An important application of MelonAntibody is its convenient use for preparing mouse polyclonal antibodies. Conventional polyclonal antibody preparation often involves rabbits, requiring more immunization shots, larger antigen doses, slower antibody production, and often outsourcing to specialized units due to high technical requirements. With MelonAntibody immunization adjuvant, any laboratory animal personnel can conveniently and rapidly prepare mouse polyclonal antibodies. A standard immunization protocol only requires immunizing five mice, resulting in 1 ml of high-quality mouse polyclonal antibodies after five weeks. (Strongly recommended for preparing mouse polyclonal antibodies using this adjuvant; 1 ml of high-quality mouse polyclonal antibodies is sufficient for the experimental needs of the vast majority of users, including applications such as ELISA, immunoblotting, flow cytometry, immunohistochemistry, and immunoprecipitation.)

Q4:Species Suitable for Adjuvants

Species suitability MelonAntibody has been immunized with various species including mice, rats, rabbits, guinea pigs, goats, alpacas, chickens, monkeys, and more, all with good results. The adjuvant itself is a mixture of multiple adjuvants, not a single adjuvant.

Q5: Increasing adjuvant dosage for low antigen concentration

For mouse immunization, it's generally recommended to use 50μl of adjuvant with 50μl of antigen. It's possible to increase the dosage slightly, or administer injections to both legs, totaling 100μl of adjuvant with 100μl of antigen. However, excessively high antigen dosages, such as over 100ug directly injected into mice, are not advisable as they may lead to strong reactions or even fatalities.

Q6: Number of mice immunized with 1ml of adjuvant

Each immunization typically requires 50μl of adjuvant for two injections, so 1ml can immunize approximately 8 mice. The standard procedure involves two injections per mouse, each with 50μl of adjuvant and 50μl of antigen, totaling 100μl per injection. While theoretically 1ml could immunize 10 mice, practical considerations such as damaged syringes or tube adhesion might limit this to around 8 mice.

Q7: Immunization and booster injection methods

The adjuvant and antigen should be mixed thoroughly immediately before immunization. Delaying immunization can cause sedimentation, resulting in uneven distribution of active components and potentially affecting immunization outcomes. For immunization, the recommended injection site for MelonAntibody water adjuvant is the inner side of the hind leg muscles of mice. For booster injections, antigens are typically injected into the abdomen at a dosage of 20-50μg in a volume of 100-200μl, avoiding the liver and bladder.

Q8: Is muscle immunization necessary?

While MelonAntibody adjuvant is optimized for muscle immunization and is primarily intended for such use, it can also be administered subcutaneously. However, intraperitoneal immunization is not recommended.

Q9: Lump formation or leakage after adjuvant use

If lumps or leakage occur, it may be due to previous intraperitoneal or subcutaneous injections, resulting in unfamiliarity with muscle immunization. Clients can try injecting deeper to prevent lump formation. Initial use should involve cleaning the hind legs with alcohol, avoiding the two inner veins, and rotating the needle after injection.

Q10: Can the immunization interval be shortened?

It's generally not recommended to shorten the immunization interval due to potential effects on antibody affinity and potency. However, for serum potency testing, it's feasible to conduct tests 7-10 days after the second immunization, provided that efficacy standards are met. Subsequent booster injections for fusion can then be performed accordingly.

Q11: Can it be used for eukaryotic expression plasmids or whole viruses?

MelonAntibody adjuvant is not suitable for plasmid immunization, but it can be used for proteins and viruses.

Q12: Can it be used for non-protein antigens such as cells or viruses?

MelonAntibody adjuvant has been validated in tens of thousands of antigen immunization experiments, showing effectiveness with a wide range of non-protein antigens including cells, viruses, peptides, polysaccharides, lipids, heavy metals, antibiotics, pesticides, drugs, food safety-related compounds, etc.

Q13: Can it be used for immunization of nucleic acid plasmids or RNA?

Nucleic acid immunization is not suitable for MelonAntibody adjuvant, and lipid nanoparticles can be considered instead.

Q14: Can it be used for vaccine development?

The primary aim of developing MelonAntibody adjuvant components was for human vaccine adjuvants, and they can be used for vaccine development. However, considering the market cost requirements for animal vaccines, clients typically consider them for use in pets or specific animals. Nevertheless, if there is an interest in cooperation to achieve cost reduction and efficiency improvement goals using high-performance water-based adjuvants, the company's capabilities in supply chain cost control and large-scale GMP production lines can likely meet those requirements.

Q15: Suboptimal efficacy after two immunizations

For antigens with weak immunogenicity or when using the product for the first time, efficacy may not improve after two immunizations. In such cases, a third immunization can be administered 14 days after the second one, typically resulting in a significant increase in efficacy. However, a maximum of four immunizations is recommended. If efficacy still doesn't improve after four immunizations, it's advisable to consider re-immunization or antigen modification, especially for small molecule antigens like peptides.

Q16: Absence of splenomegaly despite adequate antibody efficacy

This is normal. Traditional adjuvants containing BCG induce strong immune responses, leading to enlarged spleens due to the presence of many B cells targeting BCG.

Q17: Differences between adjuvants, selection criteria, and possibility of mixing

There are significant differences between rabbit and mouse adjuvants, so mixing them is not recommended. While mouse adjuvants can be mixed with each other, it's generally not advisable unless time constraints necessitate it.

Q18: Why is water-based adjuvant more expensive than Freund's adjuvant?

While the price may appear slightly higher, it saves time costs by:

1. Reducing the time needed for immunization and subsequent procedures, thus saving on mouse care expenses for a month.
2. Reducing the required antigen dosage to only 1/5-1/10 of the original amount. The cost of natural antigens can be quite high, with 1mg selling for tens of thousands of dollars.

Q20: Can fast adjuvants be used after immunization with Freund's adjuvant?

Yes, different adjuvants have similar functions, so it's possible to try using different adjuvants interchangeably.

Q21: Can it be used in parallel with aluminum adjuvants?

Yes, they can be used together. Any immunization method can generate both cellular and humoral immune responses.

Q22: Can fast adjuvants be used for booster immunization when the immunogen is cells rather than untreated protein?

The main purpose of booster immunization is to activate spleen cells to a high degree, which greatly influences the ability of hybridoma cells to secrete antibodies after fusion. Therefore, it's recommended to use the immunogen directly for booster immunization. Additionally, immunogens derived from cells often have low efficacy issues and complex compositions. The target epitope protein may not be the dominant immunogen. Previous experiences suggest that similar cell immunization approaches for antibody preparation services have generally failed to meet experimental expectations.

Q23: Species suitability

MelonAntibody has been immunized with various species including mice, rats, rabbits, guinea pigs, goats, alpacas, chickens, monkeys, and more, all with good results. The adjuvant itself is a mixture of multiple adjuvants, not a single adjuvant.