Development of Mass Spectrometry Based Techniques for the Identification and Determination of Compositional Variability in Recombinant Polyclonal Antibody Products

摘要

Recombinant polyclonal antibodies are a new class ofnprotein biologics, combining a defined number of targetspecificnantibodies, developed for therapeutic use acrossnvarious indications. Development, manufacture, and releasenof recombinant polyclonal antibodies as well characterizednbiological products have required developmentnof new chemistry, manufacturing, and control (CMC)ntechnologies. Sym001 is a recombinant polyclonal antibodynproduct containing 25 unique antibodies specific fornthe Rhesus D antigen. Sym001 drug substance is manufacturednusing a single batch technology, Sympress. Here,nwe describe the development of two novel mass spectrometrynbased methods that allows identification ofnindividual antibodies in the Sym001 drug substance,nthrough the determination of unique marker peptides ornantibody light chains. The two methods provide an unambiguousnidentification of the 25 unique antibodiesncomprised in the Sym001 drug substance. Furthermore,nthe light chain liquid chromatography-mass spectrometryn(LC-MS) method has been developed to allow the determinationnof the relative distribution of the 25 antibodies.nThe light chain LC-MS method has demonstrated linearity,nspecificity, precision, and accuracy, thus qualifying itnfor use in the quality control of recombinant polyclonalnantibodies for human use. The development of suchnquantitative methods is central for the development andnquality control of additional therapeutic recombinantnpolyclonal antibody products.