In order to produce higher quality antibodies, we can choose more suitable methods to optimize the production process. The following are some tips during the antibody production process:

Tips of Immunization Techniques

The selection of alpaca and the immune antigen are the key to the success of immunization. Choose healthy and strong, good mental state, moderate size of the alpaca is blank alpaca. The purity of the immune antigen and its correct conformation are crucial to the screening of suitable antibodies after immunizing alpacas for subsequent use, and the purity of the protein antigen is generally not less than 90%.

lymphocyte separation: Timely cell separation can effectively prevent hemolysis after blood collection to achieve the best separation effect.

The choice of immune cycle can affect the immune effect, according to experience, 1-2 weeks of immunization interval can make alpacas have a good immune response to most antigens[1].

Tips for constructing phage library

The capacity and diversity of phage library is one of the important criteria to measure the quality of the library, and the larger the capacity and the better the diversity of the library is an effective guarantee for the successful screening of nano antibodies;

The key factors affecting the capacity and diversity of the library include: degradation of RNA during RNA extraction, template and primer during reverse transcription, selection of primer and amount of template during amplification, number of PCR amplification cycles, conversion efficiency of receptive bacteria, size of linking system, etc.

Tips of Antibody screening techniques

The appropriate amount of antigen coating is related to the molecular weight of the antigen, hydrophobic and hydrophilic properties, structure, and also to the selection of coated buffer and coated medium. Reasonable coating is the basis for successful screening. If necessary, pre-experiments can be conducted to determine the conditions of coating or the method of antigen binding magnetic beads can be selected for screening[2].

The titer of the phage was determined after elution. After 2-4 rounds of antigen encapsulation by panning, the enrichment degree of the phage should be within a reasonable range.

KMD Bioscience has established a complete and mature phage antibody display technology platform. KMD Bioscience has been working on antibody research for many years. We have mature technology, with our complete antibody platform, we can offer multi-species antibody production services by using antibody phage display library technology. Meanwhile, we are devoted to providing one-stop technical services including antibody purification, antibody labeling, affinity analysis, antibody humanization, antibody affinity maturation and related services.

 

  1. De Genst E, Saerens D, Muyldermans S, Conrath K. Antibody repertoire development in camelids. Dev Comp Immunol. 2006;30(1-2):187-98.
  2. Rossotti MA, Trempe F, van Faassen H, Hussack G, Arbabi-Ghahroudi M. Isolation and Characterization of Single-Domain Antibodies from Immune Phage Display Libraries. Methods Mol Biol. 2023;2702:107-147.