Enhancement of Hybridoma Cell Growth and Anti-Hepatitis B Surface Antigen Monoclonal Antibody Production in Enriched Media with Low Serum

저혈청농축배지에서 세포성장 및 간염표면항원에 대한 단일클론항체 생산의 증가

Enhancement of hybridoma cell growth and monoclonal antibody(MAb) production by the addition of a small amount of serum into both serum-free medium and enriched medium was studied. The enriched medium was constructed by mixing a basal serum-free medium and a nutrient-fortified RPMI 1640 medium. It was supplemented with human serum albumin, insulin, transferrin, and monoethanolamine. It was found that addition of low concentration of serum with other serum-free supplements was favorable for growth of a mouse hybridoma 2c3.1 cells. The concentration of serum was determined to 0.5%. The maximum cell concentration obtained in this enriched medium supplemented with 0.5% fetal bovine serum (FBS) was $3.06{\times}10^6$ cells/ml and the concentration of secreted anti-Hepatitis surface antigen (antiHBsAg) MAb was $159.7{\mu\textrm{g}}\;/\;ml$ compared to $43{\mu\textrm{g}}\;/\;ml$ in RPMI 1640 medium with 10% FBS and $50{\mu\textrm{g}}\;/\;ml$ in previously-developed serum-free medium. The 2c3.1 cell growth and MAb production could be enhanced considerably by using the enriched medium supplemented with 0.5% FBS and serum-free supplements instead of RPMI 1640 medium or serum-free medium. The enhancement in MAb production in the enriched medium was more noticeable.

본 논문에서는 10% fetal bovine serum(FBS)를 첨가한 RPMI 1640배지에 비하여 간염바이러스 표면항원에 대한 단일클론항체 생산에 있어서 효과적은 무혈청 배지에 낮은 온도의 혈청을 첨가하여 하이브리도마_2_c3.1 세포에서 그 효과를 조사하였다, 세포성장관 단일클론항체 생산을 증가시키기 위하여 기본 무혈청배지와 RPMI 1640 배지성분들의 농도를 균형있게 강화시킨 배지를 2:1(v / v)로 혼합하여 농축배지를 조성하고, 이 배지에 2mg / ml 인혈청 알부민 $5\;{\mu\textrm{g}}\;/\;ml$ insulin, $5\;{\mu\textrm{g}}\;/\;ml$ tran-sferrin, $10\;{\mu\textrm{g}}\;/\;ml$ monoethanolmine 등의 몇가지 무혈청 첨가물들을 첨가하였다. 이 농축 배지에 fetal bovine serum(FBS)과 supplemented bovine calf serum(sBCS)을 첨가하였을때의 세포성장과 단일클론항체 생산을 비교하여 FBS농도를 변화하여 세포성장과 단일클론항체생산의 증가를 시도하였다. 0.5%를 농축배지에 첨가함으로써 세포성장과 단일클론항체생산의 증가를 보았다. 최대 세포농도는 $3.06{\times}10^6$ cells / ml이었으며, 이때 생산된 단일클론항체는 10% FBS배지의 $43.0\;{\mu\textrm{g}}\;/\;ml$ 무혈청배지의 $50\;{\mu\textrm{g}}\;/\;ml$보다 3배이상 높은 $159.7\;{\mu\textrm{g}}\;/\;ml$이 생산되었다.