Establishment of human lymphocyte cell line secreting monoclonal antibodies against Rhesus(D) antigen and sequence analysis of a human monoclonal anti-D Fab fragment / 中国病理生理杂志

ABSTRACT

AIM: To establish human lymphocyte cell line secreting monoclonal antibody against Rhesus(D) antigen and analyse the nucleotide and deduced amino acid sequences of a human monoclonal anti-D Fab fragment. METHODS: By using PCR method, the cDNA of human anti-(Rhesus D) antibody(lgM ?)Fab fragment was amplified from an Epstein- Barr-virus-transformed cell line. Cloning and subsequent sequence analysis of the Fab fragment was performed. The deduced amino acid sequence was compared and analysed with previously published sequences. RESULTS : A band of approximate 700 and 650 base pairs was amplified using lgM heavy chain primers and ? light chain primers, respectively. Sequence analysis indicated that the deduced amino acid sequences was in agreement with the characterization of the amino acid present in the human lg Fab fragment. CONCLUSION: The cloning and sequencing of a human anti-Rhesus (D) antibody Fab fragment cDNA will make benefits for production of recombinant anti-Rhesus (D) antibody and prevention of Rh haemolytic disease in the newborn.