Cloning of 3H11 mAb variable region gene and expression of 3H11 human-mouse chimeric light chain

Table 1 PCR primers for cloning variable genes of rearranged immunoglobulin light and heavy chains

5’Primers for VL Signal peptide

LL1: GGGGATATCCACCATGGAGACAGACACACTCCTGCTAT

LL2: GGGGATATCCACCATGGATTTTCAAGTGCAGATTTTCAG

LL3: GGGGATATCCACCATGGAGWCACAKWCTCAGGTCTTTRTA

LL4: GGGGATATCCACCATGKCCCCWRCTCAGYTYCTKGT

LL5: GGGGATATCCACCATGAAGTTGCCTGTTAGGCTGTTG

3’Primer for VK spanning CK and JK1,2&4

MVK: GGATACAGTTGGTGCAGTCGACTTACGTTTKATTTCCARCTT

5’Primers for VH signal peptide

VHL1: GGGGATATCCACCATGGRATGSAGCTGKGTMATSCTCTT

VHL2: GGGGATATCCACCATGRACTTCGGGYTGAGCTKGGTTTT

VHL3: GGGGATATCCACCATGGCTGTCTTGGGGCTGCTCTTCT

VHL4: GGGGATATCCACCATGATRGTGTTRAGTCTTYTGTRCCTG

3’Primer for VH spanning CH1 and JH

MVH: GACHGATGGGGSTGTYGTGCTAGCTGNRGAGACDGTGA

Table 2 Expression of chimeric light chain in transfectants determined by ELISA

ELISAsystem	Supernatant			Human Ig	Mouse Ig
	3H11	Sp2/0	Transfectant
Anti-human	0.169	0.112	1.214	1.660	0.122
Anti-mouse	1.564	0.147	0.128	0.102	1.538

a. Anti-human Fab and HRP-anti-human Fab Ab were used as catcher and second Ab. b. Anti-mouse Fab and HRP-anti-mouse Fab Ab were used as catcher and second Ab. c. ABS492.