Construction of Human Immunoglobulin Combinatorial Library and Screening of Phage Antibodies to Hepatitis B Surface
Xue Wang1, Hai-Tao Wang, Wan-Rong Chen
1Institute of Mcriology Epidemology, Academy of Military Medical Sciences, Beijing 100071, China.
Human immunoglobulin combinatorial library was generated by using phage surface-display expression system, and phage antibodies (Fab fragments) to hepatitis B surface antigen (HbsAg) were screened from it. The products by half-nested PCR using signal peptide sequences as primers were superior in quality and quantity to those by PCR with conserved sequences in the 5'-end variable regions as primers. After three round of selections by biopanning, the ratio of positive clone was 69%. The inhibition assay showed the phage antibodies to be specifically anti-HbsAg. The V(H) genes were derived from V(H) I and V(H) III, while V(L)s belonged to V(lambda) II and V(lambda) I as shown by DNA sequencing.
Human immunoglobulin combinatorial library was generated by using phage surface-display expression system, and phage antibodies (Fab fragments) to hepatitis B surface antigen (HbsAg) were screened from it. The products by half-nested PCR using signal peptide sequences as primers were superior in quality and quantity to those by PCR with conserved sequences in the 5'-end variable regions as primers. After three round of selections by biopanning, the ratio of positive clone was 69%. The inhibition assay showed the phage antibodies to be specifically anti-HbsAg. The V(H) genes were derived from V(H) I and V(H) III, while V(L)s belonged to V(lambda) II and V(lambda) I as shown by DNA sequencing.
