کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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5796759 | 1555033 | 2014 | 8 صفحه PDF | دانلود رایگان |

Single-domain variable heavy chain (VHH) antibody fragments are derived from heavy-chain antibodies of Camelids. Their comparatively small size, solubility, high affinity and specificity to the targets antigen make them suitable for many biotechnological applications. In this study, a VHH library was constructed from porcine circovirus type 2 (PCV2) vaccine immunized C. bactrianus and three VHH fragments specific to the capsid protein of PCV2 (PCV2 Cap) were selected and characterized. The selected VHH clones (VHH-c1/c3/c4) were stably expressed as soluble protein in E. coli, and were specific to PCV2 Cap except VHH-c3 which shows binding activity with both PCV1 and PCV2 Cap by ELISA. All the VHH-cs show high association rate constant and dissociation rate constant, which was 1.84 Â ÃÂ 105Â Mâ1Â sâ1, 9.00Â ÃÂ 10â3Â sâ1 for VHH-c1, 5.49Â ÃÂ 104Â Mâ1Â sâ1, 9.91Â ÃÂ 10â3Â sâ1 and 1.46Â ÃÂ 105Â Mâ1Â sâ1, 1.18Â ÃÂ 10â3Â sâ1 for VHH-c3 and VHH-c4 assessed by surface plasmon resonance (SPR). Additionally, the selected three VHH-cs can bind to different epitopes of PCV2 Cap that was determined by additive ELISA. Our study confirmed that VHHs with high affinity and specificity to PCV2 Cap can be selected from an immune VHH library, and have the potential application for effective and fast diagnostic development of PCV2.
Journal: Veterinary Immunology and Immunopathology - Volume 160, Issues 1â2, 15 July 2014, Pages 12-19