Development Of A Naïve Human Fab Antibody Library To Generate Monoclonal Antibodies Against Bmsxp Recombinant Antigen - Omar, Noorsharmimi (2015)
Antibodi adalah glykoprotein yang dihasilkan oleh sel B terhadap jasad molekul asing. Penyelidikan ini bertujuan menghasilkan antibodi Fab (fragmen ikatan antigen) manusia yang specifik terhadap antigen. Fragmen Fab terbahagi kepada satu domain malar dan domain manipulasi yang terletak pada rantai berat dan rantai ringan. Protokol optimum telah dicapai bagi menghasilkan perpustakaan Fab antibodi rekombinan daripada penderma tidak berimunisasi yang terdiri daripada tiga kumpulan etnik majoriti (Melayu, Cina dan India) di Malaysia. Kami menggunakan seluruh RNA dari limfosit periferal darah daripada 90 penderma bagi mendapatkan gen-gen manipulasi cDNA yang spesifik. Fragmen antibodi Fab yang mengekod bagi kelas IgM diamplifikasi secara individu melalui proses PCR menggunakan kepelbagaian set-set primer yang merujuk kepada bahagian manipulasi rantai berat (μ) dan rantai ringan (κ,λ). Kepelbagaian perpustakaan antibodi dicapai melalui gabungan-gabungan campuran semua rantai berat dan rantai ringan secara rawak. Antibodies are glycoproteins produced by B-cells against molecules foreign to the body. This study focused on the production of antigen specific human The Fab region consists of one constant and one variable domain which are both located at the heavy chain and the light chain. An optimized protocol for the production of a recombinant Fab library of unimmunized donors from 3 major ethnic groups (Chinese, Malay and Indian) in Malaysia was carried out. We used peripheral blood lymphocytes total ribonucleic acids (RNA) from 90 donors to obtain variable genes (V-gene) specific cDNA. The Fab-encoding regions of IgM repertoire were individually amplified by polymerase chain reaction (PCR) using a diverse set of primers corresponding to the different variable region of the heavy (μ) and the light chains (к,λ). The diversity of the library is created via random combinatorial mixing of all heavy and light V-genes.