Establishing Triple-spot MSD Assay for Manufactured Vector lot Release

Background

Our client, a Biotech company developing a unique vector containing three critical proteins all of which are transmembranes, needed support with the development and validation of a 3-spot Meso Scale Discovery (MSD) electrochemiluminescence (ECL) multiplex assay, which would be used for the Manufacturing Lot Release of their vector. The requested service was to extract three proteins from the vector, develop an MSD 3-spot assay, and transfer it back to the manufacturing site once it was validated.

The Challenge

Determining the concentrations accurately of the three proteins from unique vectors would require developing and validating a robust Lot Release MSD multiplex assay for manufacturing.  These proteins were developed by the client to modify immunogenicity. Therefore, the Sword team needed to find a wayto produce a method for extracting these proteins and putting them in a usable multiplex spot assay that their manufacturing group could use for Lot Release.  Sword Bio would utilize MSD-ECL as the system to detect all three proteins in one extraction per well.

The Solution

A highly sensitive immunoassay was developed using a unique vector extraction solution. Different capture and detection antibodies were evaluated for each protein using the MSD platform. The unique extraction solutions contained a concentration of Triton for the sample and Tween in the wash buffer. By creating the 3-spot MSD multiplex assay for the three proteins, a robust and reproducible validated assay was produced. This assay was then transferred to the client’s manufacturing team on time. The successful transfer was achieved by sending the project PI to the clients’ site to ensure the manufacturing team reproduced the data performed at the Sword’s site. Sword Bio now offers vector analysis of different proteins contained in a vector using the MSD-ECL multiplex platform for clients interested in evaluating the ratio of their multiple proteins in these unique vectors. The platform can be used to analyze vector candidates for manufacturing and in PK/PD analysis of vectors in mice, rats, or non-human primates.