Larrick J W, Coloma M J, del Valle J, Fernandez M E, Fry K E, Gavilondo-Cowley J V
Genelabs Inc., Redwood City, California.
Scand J Immunol. 1990 Aug;32(2):121-8. doi: 10.1111/j.1365-3083.1990.tb02901.x.
C6 is a potentially therapeutic murine monoclonal antibody that recognizes the class 1 outer membrane protein of Neisseria meningitidis. C6 specifically immunoblots this antigen and augments in vitro killing of N. meningitidis bacteria. We describe a general method of obtaining the heavy and light chain variable-region sequence from immunoglobulin-secreting cells. The method uses mixed polymerase chain reaction (PCR) primers designed from the 5' end of the framework 1 (FR1) sequences of the heavy and light chains, and 3'-end primers for constant-region conserved sequences. The method has been applied to the cloning and sequencing of the variable region of C6 to construct a humanized monoclonal antibody. Rapid amplification and sequencing of variable regions by this general method have multiple applications in the study of the immune response to infectious diseases.
C6是一种具有潜在治疗作用的鼠单克隆抗体,可识别脑膜炎奈瑟菌的1类外膜蛋白。C6能特异性地对该抗原进行免疫印迹,并增强体外对脑膜炎奈瑟菌的杀伤作用。我们描述了一种从免疫球蛋白分泌细胞中获取重链和轻链可变区序列的通用方法。该方法使用从重链和轻链框架1(FR1)序列的5'端设计的混合聚合酶链反应(PCR)引物,以及用于恒定区保守序列的3'端引物。该方法已应用于C6可变区的克隆和测序,以构建人源化单克隆抗体。通过这种通用方法对可变区进行快速扩增和测序在传染病免疫反应研究中有多种应用。