Generation of targeted antibody conjugates using Sortase and the AbFlex technology platform.

---

Presentation Time: 03:15 PM - 04:30 PM
Poster Board Number: B932
Abstract ID: 5817

Presenting Author:
Mary Anne Jelinek , Associate Director, R&D , Active Motif

Abstract:

The miniaturization of experiments to the single cell scale and a growing interest in multiplexing analyte detection is a major driver for the development of novel antibody conjugation technologies that are robust, and which also preserve antibody function.  Relative to two step detection modalities, direct primary antibody conjugates provide enhanced specificity and selectivity. We have developed the AbFlex platform, a versatile epitope tagging system for the carboxyl terminus of recombinant mouse and rabbit immunoglobulin heavy chains. It consists of a tandem array of three tags: 1) The Sortase tag recognition sequence (LPXTG); 2) The motif recognized by the BirA biotin ligase, followed by 3) the 6XHis Tag, which can be used with nickel-based purification systems.

Sortase A belongs to a family of transpeptidases found in Gram-positive bacteria that catalyze the attachment of poly-Glycine labels to the Sortase recognition sequence under physiological conditions and with reaction times of approximately one hour. We describe the use of the Staphylococcus aureus Sortase A5 pentamutant that is significantly more active than wild-type Sortase to provide a faster, more efficient labeling reaction. Performance of various recombinant mouse and rabbit antibody conjugates in a variety of applications will be presented.