The Easy Guide To Custom Antibody Production
Antibodies are protein molecules produced in the body during an immune response to antigens of foreign molecules attacking the body. Besides their immune function, another distinguishing antibody characteristic is their high specificity to antigens.
Consequently, antibody technology is crucial in multiple biotechnology disciplines, including protein research, diagnostics, assay tests, and vaccine development. Antibody production services use expression systems to generate antibodies for various projects.
However, advances in biotech make it possible to produce custom antibodies from unique peptides or proteins provided by a customer for their project. Below are the steps involved in custom antibody production.
Purify Target Antigens
As stated above, immune cells automatically produce antibodies corresponding to a specific antigen when the antigen (pathogen) enters the body. Therefore, recombinant custom antibody production utilizes the same principle to generate custom antibodies in vitro.
The process begins with the acquisition of an antigen matching the target antibody. Antigens could be any portion of a pathogen like bacteria, fungi, viruses, or toxins; therefore, peptides, polypeptides, or protein fragments from such pathogens are acceptable antigens fragments.
However, most custom antibody production services stipulate the minimum antigen quantity ideal for custom antibody generation. Moreover, you can begin the custom antibody production process with antigen design if the target antigen does not occur naturally.
According to one literature review, the purity of a sample antigen influences the generation of the target antibody; a higher antigen purity translates to high-quality, high-throughput target antibodies. The review further states that the rule of thumb in antigen purification is that the antigen sample should contain no more than 1%-2% of contaminants to avoid non-targeted immune responses.
Custom antibody production companies use different antigen purification methods to separate the target antigen from a complex mixture. The purification techniques include isoelectric focusing, chromatofocusing, affinity chromatography, and high-performance liquid chromatography. Each antigen purification method has advantages and disadvantages, so consult the experts on the ideal purification method for your project.
Immunogenic Carrier Protein Selection
In most cases, the purified antigen is not large enough to link to T-cell receptors and trigger an immune response. Therefore, antibody production services use carrier proteins to enlarge the ideal molecular size to trigger an immune response. The process entails coupling or conjugating minute antigen molecules like peptides or haptens with select molecules to form a large, complex molecule.
However, different proteins have unique characteristics, meaning not all protein molecules make ideal carrier proteins. Therefore, consider the following tips when choosing a carrier protein.
First, consider using larger carrier protein molecules to couple with the purified antigen because larger molecules have the elements necessary to activate T-cells. Second, consider conjugating the purified antigen with a protein molecule genetically distinct from the host or expression system to be used in the target antibody production.
The host’s T-cells may fail to view a genetically similar protein as a threat. Lastly, consider using a highly soluble protein molecule to make conjugation with the purified protein easier.
Conjugate An Immunogen/ Immunogen Design
An immunogen is a molecule or substance that triggers a cell-mediated immune response. Since the purified antigen lacks immunogenic properties, you must conjugate it with a carrier protein to form an immunogen to be used to trigger an immune response.
Conjugate vaccines are good immunogens that merge a purified antigen with viable carrier proteins. The host animal’s immune cells recognize the antigen fragment in the immunogen, releasing corresponding antibodies. However, only conjugate the immunogen after finding a compatible carrier protein.
Immunize The Animal Host
Immunization entails the active exposure of a host to the antigen to generate target antibodies. The conjugate immunogen is the antigen that triggers an immune response in the host animal’s cells during custom antibody production.
However, the custom antibody production company must prepare the immunogen for immunization using an adjuvant. An adjuvant is a molecule, compound, or solution that lengthens the immune response’s durations and boosts the antibodies’ potency while minimizing or hampering toxicity. The selection of the ideal adjuvant depends on the purified antigen’s physical and chemical properties.
Second, a vaccine schedule is a prerequisite for the immunization process. The immunization schedule allows the scientists conducting the custom antibody production process to monitor the vaccine’s effects at different doses. Subsequent vaccinations also increase the target antibody levels against the antigen. The number of scheduled vaccines varies depending on the host animal and antibody type; hence the antibody production companies offer various protocol packages.
Screen Host Serum For Antibody Titers Or Serum
The antibodies produced in the host animal are present in its blood serum. Therefore, screening is necessary to measure the target antibody’s presence and level (titer). Screening is necessary before bleeding to collect the serum with the target antibodies.
Antibody Purification
Antibody purification entails isolating the retrieved antibodies from the animal host serum, in the case of polyclonal antibodies, and the hybridoma cell from the serum for monoclonal antibodies.
Custom antibody production services use standard protein purification methods, including ion exchange chromatography, gel filtration chromatography, and hydrophobic interaction chromatography. Each method has advantages and disadvantages, depending on the target antibody’s characteristics.
Antibody Fragmentation
Antibody fragmentation entails using reducing agents and enzymes and is the stage at which the bulk of antibody customization occurs. One literature review shows that antibody fragmentation alters the antibody molecules’ physiochemical properties.
Conclusion
Custom antibody production is essential for advancing research in medicine and therapeutics. Peptide synthesis companies offer different custom peptide production services, so learning the basics of the process is crucial in choosing the ideal package for your project.
