Use of bispecific antibodies in molecular velcro assays whose specificity approaches the theoretical limit of immunodetection for Bordetella pertussis

Clinical and Diagnostic Laboratory Immunology
X L TangM R Suresh

Abstract

A bispecific monoclonal antibody (bsMAb) that detects Bordetella pertussis, the causative agent of whooping cough, and horseradish peroxidase (HRPO) has been developed by use of the quadroma technology. A quadroma, P123, was produced by fusing two well-characterized hybridomas against the bacterium and the enzyme and was subcloned to obtain a stable bsMAb-secreting cell line. The quadroma was theoretically expected to produce up to 10 different molecular species of immunoglobulins, so secreted bispecific antibody was complexed with excess HRPO and the HRPO-bsMAb complex was purified in one step by benzhydroxamic acid-agarose affinity cochromatography. An ultrasensitive homosandwich molecular "velcro" enzyme-linked immunosorbent assay for the detection of B. pertussis whole bacteria with HRPO-bsMAb was established in both microplate and nasopharyngeal swab formats. This assay demonstrates a high sensitivity that approaches the theoretical limit of detection of one bacterium. This new nanoprobe can be used to develop a new generation of assays that are simple, inexpensive alternatives to quantitative PCR and that can be used by clinical laboratories. This strategy of homosandwich assays with solid-phase monospecific antibodies an...Continue Reading

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Citations

Oct 8, 2013·Trends in Biotechnology·Hannah ByrneRichard J O'Kennedy
Nov 12, 2013·Diagnostic Microbiology and Infectious Disease·Advaita GangulyHoon H Sunwoo
Jul 16, 2008·Journal of Virological Methods·Sriram KammilaMavanur R Suresh
Oct 6, 2005·Journal of Immunological Methods·Welson W WangMavanur R Suresh
Apr 15, 2015·Journal of Virological Methods·Advaita GangulyHoon H Sunwoo
Dec 12, 2018·International Journal of Biological Macromolecules·Jyoti GuptaM Saleemuddin
May 25, 2021·Frontiers in Immunology·Jiabing MaCheng Qian

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