A whole-molecule immunocapture LC-MS approach for the in vivo quantitation of biotherapeutics.
OBJECTIVE
Large-molecule biotherapeutic quantitation in vivo by LC-MS has traditionally relied on enzymatic digestion followed by quantitation of a 'surrogate peptide' to infer whole-molecule concentration. MS methods presented here measure the whole molecule and provide a platform to better understand the various circulating drug forms by allowing for variant quantitation.


RESULTS
An immunocapture LC-MS method for quantitation of a biotherapeutic monoclonal antibody from human plasma is presented. Sensitivity, precision and accuracy for each molecular portion are presented along with an example of glycoform variant quantitation.


CONCLUSIONS
The method is presented as a basic platform to be further developed for Good Practice (GxP) applications, critical quality attribute analysis or general understanding of molecular forms present as required for the wide range of drug development processes.
Bioanalysis, Immunogenicity & Biomarkers, In vitro/In vivo Translation Platform, R&D Platform Technology & Science, GSK, 709 Swedeland Road, King of Prussia, PA 19406, USA.
John F
JF
Kellie
Bioanalysis, Immunogenicity & Biomarkers, In vitro/In vivo Translation Platform, R&D Platform Technology & Science, GSK, 709 Swedeland Road, King of Prussia, PA 19406, USA.
Jonathan R
JR
Kehler
Bioanalysis, Immunogenicity & Biomarkers, In vitro/In vivo Translation Platform, R&D Platform Technology & Science, GSK, 709 Swedeland Road, King of Prussia, PA 19406, USA.
Thomas J
TJ
Mencken
Bioanalysis, Immunogenicity & Biomarkers, In vitro/In vivo Translation Platform, R&D Platform Technology & Science, GSK, Park Road, Ware, Hertfordshire, SG12 0DP, UK.
Richard J
RJ
Snell
Bioanalysis, Immunogenicity & Biomarkers, In vitro/In vivo Translation Platform, R&D Platform Technology & Science, GSK, 709 Swedeland Road, King of Prussia, PA 19406, USA.
Charles S
CS
Hottenstein
D000911:Antibodies, Monoclonal / Q000097:blood / Q000276:immunology*; D001774:Blood Chemical Analysis / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D006031:Glycosylation; D006801:Humans; D007118:Immunoassay; D010455:Peptides / Q000097:blood* / Q000276:immunology; D053719:Tandem Mass Spectrometry
D000911:Antibodies, Monoclonal; D010455:Peptides
biotherapeutic quantitation; immunoassay LC–MS; immunocapture LC–MS; intact protein quantitation; top-down MS; whole-molecule MS