Label-free and amplified quantitation of proteins in complex mixtures using diffractive optics technology.
Immunoaffinity approaches remain invaluable tools for characterization and quantitation of biopolymers. Their application in separation science is often limited due to the challenges of immunoassay development. Typical end-point immunoassays require time consuming and labor-intensive approaches for optimization. Real-time label-free analysis using diffractive optics technology (dot) helps guide a very effective iterative process for rapid immunoassay development. Both label-free and amplified approaches can be used throughout feasibility testing and ultimately in the final assay, providing a robust platform for biopolymer analysis over a very broad dynamic range. We demonstrate the use of dot in rapidly developing assays for quantitating (1) human IgG in complex media, (2) a fusion protein in production media and (3) protein A contamination in purified immunoglobulin preparations.
Cleverley
Steve
S
TouchDown Biomarketing BV, Bemmel, The Netherlands.
Chen
Irene
I
Houle
Jean-François
JF
D001704:Biopolymers / Q000032:analysis; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods*; D002138:Calibration; D045424:Complex Mixtures / Q000737:chemistry; D004867:Equipment Design; D017743:Equipment Reuse; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods*; D007074:Immunoglobulin G / Q000032:analysis; D055095:Optics and Photonics / Q000295:instrumentation* / Q000379:methods*; D011485:Protein Binding; D011506:Proteins / Q000032:analysis*; D011993:Recombinant Fusion Proteins / Q000032:analysis; D013205:Staphylococcal Protein A / Q000032:analysis
D001704:Biopolymers; D045424:Complex Mixtures; D007074:Immunoglobulin G; D011506:Proteins; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A