title: Development of mass spectrometry based techniques for the identification and determination of compositional variability in recombinant polyclonal antibody products.
abstract: Recombinant polyclonal antibodies are a new class of protein biologics, combining a defined number of target-specific antibodies, developed for therapeutic use across various indications. Development, manufacture, and release of recombinant polyclonal antibodies as well characterized biological products have required development of new chemistry, manufacturing, and control (CMC) technologies. Sym001 is a recombinant polyclonal antibody product containing 25 unique antibodies specific for the Rhesus D antigen. Sym001 drug substance is manufactured using a single batch technology, Sympress. Here, we describe the development of two novel mass spectrometry based methods that allows identification of individual antibodies in the Sym001 drug substance, through the determination of unique marker peptides or antibody light chains. The two methods provide an unambiguous identification of the 25 unique antibodies comprised in the Sym001 drug substance. Furthermore, the light chain liquid chromatography-mass spectrometry (LC-MS) method has been developed to allow the determination of the relative distribution of the 25 antibodies. The light chain LC-MS method has demonstrated linearity, specificity, precision, and accuracy, thus qualifying it for use in the quality control of recombinant polyclonal antibodies for human use. The development of such quantitative methods is central for the development and quality control of additional therapeutic recombinant polyclonal antibody products.
authors.affiliation: Swedish Orphan Biovitrum AB, Tomtebodavagen 23 A, SE-112 76 Stockholm, Sweden.
authors.forename: Pia
authors.initials: P
authors.lastname: Persson
authors.forename: Anders
authors.initials: A
authors.lastname: Engström
authors.forename: Lone K
authors.initials: LK
authors.lastname: Rasmussen
authors.forename: Erland
authors.initials: E
authors.lastname: Holmberg
authors.forename: Torben P
authors.initials: TP
authors.lastname: Frandsen
mesh_terms: D000906:Antibodies / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000032:analysis; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012204:Rh-Hr Blood-Group System / Q000276:immunology; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D000906:Antibodies; D007147:Immunoglobulin Light Chains; D011994:Recombinant Proteins; D012204:Rh-Hr Blood-Group System; C026410:Rho(D) antigen