Abstract
The C2 domain of streptococcal protein G is a small (55 residue) peptide with immunoglobulin-binding activity. Following codon optimization, the gene was divided into four oligonucleotide fragments and amplified by overlap PCR. The recombinant plasmid pET30a-C2 was transformed into Escherichia coli Rosetta (DE3) PLysS for expression. After purification by Ni–NTA, the fusion protein was identified by western-blotting, Dot-ELISA and ELISA. His-tagged C2 bound to human, rabbit, cattle, pig, goat, mouse or guinea pig IgG had no affinity for goose, duck, wild duck, wild turkey and red-crowned crane IgY. Its affinity for chicken IgY, however, was comparable to that of guinea pig IgG. The C2 domain may therefore provide an ideal material for the purification and detection of immunoglobulin G from various mammals.
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Acknowledgments
This study was supported by grants from the Earmarked Fund for China Agriculture Research System (No. CARS-37), the sub-project of National 12th 5-year Support Key Projects (2012BAD12B03, 2012BAD12B05) and the Key Technologies Research and Development Program of Heilongjiang province (GA09B302). We would like to express our sincere appreciation to the anonymous reviewers for their insightful comments, which have greatly aided us in improving the quality of the paper.
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Cao, Y., Li, D., Cao, C. et al. Characterization of the optimized C2 domain of protein G: finding its additional chicken IgY-binding ability. Biotechnol Lett 35, 1441–1447 (2013). https://doi.org/10.1007/s10529-013-1221-7
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DOI: https://doi.org/10.1007/s10529-013-1221-7