Binding and neutralization characteristics of a panel of monoclonal antibodies to human papillomavirus 58

Abstract

Human papillomavirus (HPV) 58 is a high-risk HPV type associated with progression to invasive genital carcinomas. We developed six monoclonal antibodies (mAbs) against HPV58 L1 virus-like particles that bind conformational epitopes on HPV58. The hybridoma cell lines were adapted to serum- and animal component-free conditions and the mAb supernatants were affinity-purified. The six mAbs neutralized HPV58 pseudoviruses (PsVs) and 'quasivirions' with different capacities. The mAbs differed in their ability to prevent PsV58 attachment to HaCaT cells, to the extracellular matrix (ECM) deposited by HaCaT cells, to heparin and to purified human laminin 5, a protein in the ECM. These mAbs provide a unique set of tools to study the binding properties of a previously untested, high-risk HPV type and the opportunity to compare these characteristics with the binding of other HPV types.

Fig. 1.

(a) Binding titres of mAbs to HPV58 PsVs, (b) neutralization of PsV58 and (c) mAb inhibition of binding of PsVs to heparin–BSA or LN5 by ELISA-based binding or neutralization assays. One asterisk indicates a P-value of <0.001 and two asterisks indicate a P-value of <0.01. The absorbance is indicated on the y-axis and the dilution of antibody (a, b) or well treatment (c) is indicated on the x-axis. Each figure is representative of three separate experiments.

Fig. 2.

mAb inhibition of PsV58 binding to HaCaT cells or HaCaT-derived ECM. (a) PsV58 binding to cells; (b) PsV58 binding to ECM; (c) merge of PsV58 and LN5 staining. This figure is representative of three separate experiments. Bars, 10 μm.