公开(公告)号：KR1020000040714A

公开(公告)日：2000-07-05

申请号：KR1019980056432

申请日：1998-12-19

Applicant: 한국과학기술원

Inventor： 이균민 ,  류준수

IPC: C12N1/00

Abstract: PURPOSE: Production method of monoclonal antibody by fed batch culture using low osmotic pressure medium as a starting medium and feeding concentrate of the starting medium is provided which relieves increment of osmotic pressure resulted from added medium concentrate, prolongs the life span of cells and consequently, produces high density of monoclonal antibody. CONSTITUTION: Starting medium, having low osmotic pressure about 200mOsm/kg - 250mOsm/kg, and medium concentrate, from which salts are removed completely, are used to produce monoclonal antibody from hybridoma cells. Starter medium is prepared from just removing NaCl of DMEM medium. Medium concentrate is prepared from DMEM medium after removing NaCl, KCl, calcium chloride, sodium hydrocarbonate, sodium hydrogenphosphate, magnesium sulfate and ferric nitrate without serum. The salt free medium is concentrated 5-10 times. Medium concentrate is added when cell reaches stationary phase and the frequency of feeding is 3-4 times every 24 or 48 hour. ELISA analyses reveal that production of monoclonal antibody with low osmotic pressure medium is increased 6 times in batch culture and 1.7 times in feed batch culture.

Abstract translation: 目的：提供使用低渗透压培养基作为起始培养基的饲料分批培养和起始培养基的进料浓缩物的单克隆抗体的生产方法，其减轻了由添加的培养基浓缩物引起的渗透压增加，延长了细胞的寿命， 产生高密度的单克隆抗体。 构成：使用具有约200mOsm / kg-250mOsm / kg的低渗透压的起始培养基和从其中完全除去盐的中浓缩物用于从杂交瘤细胞产生单克隆抗体。 起始介质是从除去DMEM培养基的NaCl制备的。 在去除NaCl，KCl，氯化钙，碳酸氢钠，磷酸氢钠，硫酸镁，无血清的硝酸铁之后，从DMEM培养基中制备中浓缩物。 无盐培养基浓缩5-10倍。 当细胞达到固定相时加入中浓缩物，每24或48小时喂食次数为3-4次。 ELISA分析显示，在分批培养中生产具有低渗透压培养基的单克隆抗体增加6倍，饲料分批培养中增加1.7倍。