Novel approach for rapid generation of human antibodies to RNA-modifications

Period of Performance: 07/01/2016 - 06/30/2017

$225K

Phase 1 SBIR

Recipient Firm

Abzyme Therapeutics, LLC
POTTSTOWN, PA 19464
Principal Investigator

Abstract

Project Summary: It is known that protein activity can be modulated by post-translational proteinphosphorylation. Phospho-specific antibodies have greatly advanced understanding of the mechanisms ofprotein activation and signaling in cellular pathways. Post-transcriptional RNA modification is morecomplicated as there are 66 known RNA modifications that occur in thousands of RNAs in eukaryotic cells. Anumber of these RNA modifications have been shown to influence the development and physiology of thecentral nervous system (CNS). The availability of antibodies against RNA modifications at specific RNAsequences analogous to phospho-specific antibodies will facilitate investigation of the relationships betweenspecific RNA modifications and their cellular function. Unfortunately due to in vivo instability of RNAs and theirnon-immunogenicity, it is difficult to raise antibodies against RNA modifications in animals. The goal of thisproject is to develop an in vitro novel platform for rapid generation and robust production of a set of antibodiesagainst specific RNA modifications. As proof-of-principle, in Phase I, a yeast-based self-diversifying library ofhuman single-chain variable fragment (ScFv) will be established and ScFv antibodies capable of recognizingInosine (I), 5-methylcytosine (m5C), N6-methyladenosine (m6A) nucleosides and mRNA containing Adenine-to-Inosine (A-to-I) modification at the site A of mouse 2C receptor will be developed. In phase II, the effort willbe dedicated to develop antibodies capable of recognizing RNA modification at a given RNA context such asA-to-I modification of A, B, C, D and E sites of mouse 2C receptor mRNA as well as A-to-I modifications foundin other mRNA such as GluR2 and KA receptors. In addition, the developed platform will be used forgenerating antibodies to other RNA modifications. Availability of such reagents will enable researchers tomonitor the effect of specific RNA modification on the structure or function of respective individual RNA or toanalyze tissue sections, e.g. tumor samples, for the occurrence of such modifications.