High affinity mouse-human chimeric Fab against Hepatitis B surface antigen

Table 1 Oligonucleotide primers used for generation of the chimeric Fd and light chain

Primer	Sequence
5’ primer for VH (5H23M)	5’- AG GTC CAG CTT CTC GAG CCC GGG GC -3’
3’ primer for VH (Fd3)	5’-CGA TGG GCC CTT GGT GGA GGC TGA AGA GAC AGT GAC TGA GGT TCC-3’
5’ primer for CH1 of human IgG1 (Fd5)	5’-GGA ACC TCA GTC ACT GTC TCT TCA GCC TCC ACC AAG GGC CCA TCG-3’
3’ primer for CH1 of human IgG1 (CG1Z)	5’- GCA TGT ACT AGT TTT GTC ACA AGA TTT GGG -3’
5’ primer for VL (5L35)	5’-CCA GAT GTG AGC TCG TGA TGA CCC AGA CTC CA-3’
3’ primer for VL (K3)	5’-CAG ATG GTG CAG CCA CAG TCC GTT TGA GTT CCA GCT TGG-3’
5’ primer for CL of human κ chain (K5)	5’-CCA AGC TGG AAC TCA AAC GGA CTG TGG CTG CAC CAT CTG-3’
3’ primer for CL of human κ chain (Ck1d)	5’- GCG CCG TCT AGA ATT AAC ACT CTC CCC TGT TGA AGC TCT TTG TGA CGG GCG AAC TCA G -3’

Primer names are given in parenthesis. Sites for restriction enzymes are shown as underlined. Complementary overhangs are shown in bold letters.

Table 2 Yields of the anti-HBs chimeric Fab in different E. coli strains

E. coli strain	Yield (μg/L)1
XL1-Blue	40
AD494	62
BL21 Codon Plus	40
TG1	8

¹Estimated by Bradford assay, after purification by protein G column