CONSTRUCTION OF PHAGE LIBRARY OF HUMAN ANTIBODY FROM GLIOMA PATIENTS / 解放军医学杂志

ABSTRACT

To construct human antibody repertory with phage display technology. Total RNA was extracted from peripheral blood lymphocytes (PBL) of clinical glioma patients and subjected to reverse transcription PCR. Variable region genes(V ? and V H genes) were amplified from the cDNA using nested polymerase chain reaction. V ? gene library was first constructed and attached to the Linker which could specially connect V ? gene and V H gene, then V H gene was cloned into V ? gene library to build ScFv repertoire. Positive colonies were selected randomly and sequenced. The results implied that amplified human V ? gene and V H gene could be derived from PBL of clinical glioma patients. ScFv phage antibody gene repertoire in about 2?10 6 capacity was successfully constructed. The sequencing indicated that the cloned genes encoded variable regions of heavy chains of human antibody and were highly homologous with human kappa heavy chain group III of Kabat's database. Then human antibody repertoire was successfully constructed with phage display technology. The present study can be used as a foundation for succeeding screening of specific antibody against human glioma.