Bong Ji-Hong, Kim Tae-Hun, Jung Jaeyong, Lee Soo Jeong, Sung Jeong Soo, Lee Chang Kyu, Kang Min-Jung, Kim Hyun Ok, Pyun Jae-Chul
Department of Materials Science and Engineering, Yonsei University, 50 Yonsei-Ro, Seodaemun-Gu, Seoul, 03722 Republic of Korea.
Molecular Recognition Research Center, Korea Institute of Science and Technology (KIST), Seoul, 02792 Republic of Korea.
Biochip J. 2020;14(4):358-368. doi: 10.1007/s13206-020-4404-z. Epub 2020 Oct 22.
Anti-coronavirusdisease-2019 (COVID-19; anti-severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2)) antibodies against nucleoprotein (NP) were purified from pig sera. Following the separation of the antibody fraction using a protein-A column, the final yield of the purified antibodies against SARS-CoV-2 NPs was estimated to be 0.26 ± 0.05 % (absolute amount of 143.4 ± 25.2 ng, n=5) from 1 mL of pig sera. The binding activities of the isolated antibodies were confirmed using immunoassay and immunostaining. Based on the specific binding activity to NPs, a quantitative assay was performed using a surface plasmon resonance (SPR) biosensor. From the doseresponse curve, the binding constant (Kd) was calculated to be 185 pM and the limit of detection was estimated to be 1.02 pM. The SPR biosensor with the isolated antibodies against SARS-CoV-2 NPs was applied for the detection of SARS-CoV-2, MERS-CoV, and CoV strain 229E in culture fluid.
抗2019冠状病毒病（COVID-19；抗严重急性呼吸综合征冠状病毒2（SARS-CoV-2））核蛋白（NP）抗体从猪血清中纯化。使用蛋白A柱分离抗体组分后，从1 mL猪血清中纯化的抗SARS-CoV-2 NP抗体的最终产量估计为0.26±0.05%（绝对量为143.4±25.2 ng，n=5）。使用免疫测定和免疫染色确认分离抗体的结合活性。基于对NP的特异性结合活性，使用表面等离子体共振（SPR）生物传感器进行定量测定。从剂量反应曲线计算出结合常数（Kd）为185 pM，检测限估计为1.02 pM。将带有分离的抗SARS-CoV-2 NP抗体的SPR生物传感器应用于检测培养液中的SARS-CoV-2、中东呼吸综合征冠状病毒（MERS-CoV）和229E冠状病毒株。
