Stability of anti-N clone 1391G2 antibody secreting cell line

Abstract

Background: Cell line secreted anti-N (1391G2) is a murine monoclonal hybridoma cell that Antiserum and standard cell preparation section, National Blood Centre, Thai Red Cross Society used for culture and expand in large scale for harvest supernatant to produce blood group reagent anti-N for distributing to blood bank and hospital in Thailand. Therefore, this cell line must be regularly tested for stability of antibody secreting.

Objectives: This study aimed to maintained and select perfectly cell line secreted anti-N (1391G2) 3 generations.

Methods: Cell line secreted anti-N 1st generation namely 1391G2 was thawed from liquid nitrogen and selected the best single cell line with character of good growth rate and high titer antibody secreting using limiting dilution method. After that, selected cell line will be frozen in liquid nitrogen for 7 days. In addition, the 1st generation cell line that separated from freezing was cultured for harvest supernatant to serology test. The 2ndgeneration cell line was thawed after frozen for 7 days for limiting dilution method again until 3rd generation. Serology test of supernatant from all generation cell line consisted of antibody identification with panel cell and antibody titration with red blood cell group O that has M-N+ and M+N+ antigen.

Results: Anti-N cell line 3 generations stably expressed anti-N which antibody titer with M-N+ and M+N+ were 128 and 64, respectively.

Conclusion: All generation of anti-N titer was comparable to American Association of Blood Banks (AABB) standard. This result validates the quality of anti-N production for blood typing reagent from Nation blood center, Thai red cross society to ensure safety of blood donation and transfer process.