Aim: To explore the conditions of solid-phase screening phage antibody library and to provide the experimental basis for the design of screening project.
Methods: Diverse antibodies including HEV NE2-specific and non-specific humanized phage antibodies were used to study the screening conditions, such as the binding time of phage antibodies to antigen, the concentration of coating antigen, the washing times and elution method.
Results: The best binding time of positive phage antibody to antigen was 1 min. The highest positive rate of screening was obtained under the conditions of washing for 20 to 30 times and pH value of the washing solution being 5. The concentration of the coating antigen had no obvious influence on the positive rate of screening. Higher positive rate was obtained by using 10 mg/L antigen to competitively elute for 1 h.
Conclusion: Solid-phase screening of the phage antibody library is a very complex process, in which there are close relationship between the conditions, therefore, appropriate readjustment should be made for screening conditions according to concrete conditions.