| Info. | Vol.15 - No.4 (2021.12.20) |
|---|---|
| Title | Isolation of Antibodies Against the Spike Protein of SARS-CoV from Pig Serum for Competitive Immunoassay |
| Authors | Jaeyong Jung1, Ji-Hong Bong1, Tae-Hun Kim1, Jeong Soo Sung1, Changkyu Lee1, Min-Jung Kang2, Hyun Ok Kim3, Hyun-Jin Shin4 & Jae-Chul Pyun1,* |
| Institutions | 1Department of Materials Science and Engineering, Yonsei University, 50 Yonsei-Ro, Seodaemun-Gu, Seoul 03722, Republic of Korea 2Molecular Recognition Research Center, Korea Institute of Science and Technology (KIST), Seoul 02792, Korea 3Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul 03722, Republic of Korea 4College of Veterinary Medicine, Chungnam National University, Daejeon, Korea |
| Abstract | Several endemic corona viruses (eCoVs) have been reported to be the most common etiologic agents for the seasonal common cold and also cause pneumonia. These eCoVs share extensive sequence homology with SARS-CoV-2, and immune responses to eCoVs can cross-react with SARS-CoV-2 antigens. Based on such cross-reactivity of antigens among eCoVs, the IgG antibodies against the spike protein (SP) of severe acute respiratory syndrome coronavirus (SARS-CoV) were isolated from pig serum using magnetic beads immobilized with SARS-CoV SP and a protein-A column. The selectivity of the isolated antibodies was tested using different types of antigens, such as SARS-CoV-2 nucleoprotein (NP), influenza A virus (Beijing type), influenza B virus (Tokio and Florida types), human hepatitis B virus surface antigen (HBsAg), and bovine serum albumin (BSA). From the selectivity test, the anti-SP antibodies isolated from pig serum had sufficient selectivity to other kinds of viral antigens, and the apparent binding constant of the isolated antibodies was approximately 1.5 × 10 –8 M from the surface plasmon resonance (SPR) measurements. Finally, the isolated anti-SP antibodies were applied to the immunoassay of SP using competitive immunoassay configuration. The feasibility of the detection as well as the quantitative analysis of the SARS-CoV viral culture fluid was determined using four viral culture samples, namely, SARS-CoV, SARS-CoV-2, MERS-CoV, and CoV-229E. |
| Keyword | SARS-CoV spike protein, Competitive immunoassay, Antibody isolation, Pig serum |
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