Quantifying Neutralizing Antibodies in Patients with COVID-19 by a Two-Variable Generalized Additive Model

Kuan Ting Liu, Yu Nong Gong, Chung Guei Huang, Peng Nien Huang, Kar Yee Yu, Hou Chen Lee, Sun Che Lee, Huan Jung Chiang, Yu An Kung, Yueh Te Lin, Mei Jen Hsiao, Po Wei Huang, Sheng Yu Huang, Hsin Tai Wu, Chih Ching Wu, Rei Lin Kuo, Kuan Fu Chen, Chuan Tien Hung, Kasopefoluwa Y. Oguntuyo, Christian S. StevensShreyas Kowdle, Hsin Ping Chiu, Benhur Lee, Guang Wu Chen, Shin Ru Shih

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


Considering the urgent demand for faster methods to quantify neutralizing antibody titers in patients with coronavirus (CoV) disease 2019 (COVID-19), developing an analytical model or method to replace the conventional virus neutralization test (NT) is essential. Moreover, a "COVID-19 immunity passport"is currently being proposed as a certification for people who travel internationally. Therefore, an enzyme-linked immunosorbent assay (ELISA) was designed to detect severe acute respiratory syndrome CoV 2 (SARS-CoV-2)-neutralizing antibodies in serum, which is based on the binding affinity of SARS-CoV-2 viral spike protein 1 (S1) and the viral spike protein receptor-binding domain (RBD) to antibodies. The RBD is considered the major binding region of neutralizing antibodies. Furthermore, S1 covers the RBD and several other regions, which are also important for neutralizing antibody binding. In this study, we assessed 144 clinical specimens, including those from patients with PCR-confirmed SARS-CoV-2 infections and healthy donors, using both the NT and ELISA. The ELISA results analyzed by spline regression and the two-variable generalized additive model precisely reflected the NT value, and the correlation between predicted and actual NT values was as high as 0.917. Therefore, our method serves as a surrogate to quantify neutralizing antibody titer. The analytic method and platform used in this study present a new perspective for serological testing of SARSCoV- 2 infection and have clinical potential to assess vaccine efficacy.

Original languageEnglish
Article numbere00883-21
Issue number1
StatePublished - Feb 2022


  • SARS-CoV-2
  • enzyme-linked immunosorbent assay
  • neutralizing antibody
  • receptor-binding domain
  • spike protein
  • two-variable generalized additive model


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