Christian R B, Couto J R, Peterson J A, Ceriani R L
Cancer Research Fund of Contra Costa, Walnut Creek, California 94596, USA.
Hybridoma. 1996 Apr;15(2):155-8. doi: 10.1089/hyb.1996.15.155.
Mc5, a murine monoclonal antibody that binds to human breast epithelial mucin (BEM), has been shown to be a promising reagent in the diagnosis of breast cancer. We have cloned cDNAs encoding both variable regions of Mc5 (VL and VH) as well as the CL and CH1 constant regions. Mc5 is an IgG1, kappa antibody. We have constructed an IgG1, kappa human/mouse chimeric antibody (by inserting the murine VH and VL-encoding cDNAs into plasmids encoding human constant domains), and expressed it in SP2/0-Ag14 mouse myeloma cells. The affinity of chimeric Mc5 (chMc5) for BEM is 4.4 x 10(8) M-1. Mc5 binds BEM with an affinity constant of 2.8 x 10(8) M-1. Purified chMc5 and purified Mc5 gave similar competition curves when tested against either 125I-labeled Mc5 or 125I-labeled chMc5 for binding to BEM in a competition radioimmunodetection format. Additionally, chMc5 used in breast carcinoma tissue staining stained as well as the original Mc5.
Mc5是一种与人类乳腺上皮粘蛋白（BEM）结合的鼠单克隆抗体，已被证明是诊断乳腺癌的一种有前景的试剂。我们已经克隆了编码Mc5两个可变区（VL和VH）以及CL和CH1恒定区的cDNA。Mc5是一种IgG1 κ抗体。我们构建了一种IgG1 κ人/鼠嵌合抗体（通过将编码鼠VH和VL的cDNA插入编码人恒定结构域的质粒中），并在SP2/0-Ag14小鼠骨髓瘤细胞中表达。嵌合Mc5（chMc5）对BEM的亲和力为4.4×10⁸ M⁻¹。Mc5以2.8×10⁸ M⁻¹的亲和常数结合BEM。在竞争放射免疫检测形式中，当用¹²⁵I标记的Mc5或¹²⁵I标记的chMc5检测结合BEM时，纯化的chMc5和纯化的Mc5给出了相似的竞争曲线。此外，用于乳腺癌组织染色的chMc5与原始Mc5染色效果相同。
