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Assessing Effects of Freeze-Thaw on Biotinylated Macromolecules Using Gyrolab™
Author(s): Xiaodong F Liu, Roni Weaver, Charles Hottenstein, Matthew Szapacs, Lee Abberley, LingSing Chen and Xun WangXiaodong F Liu, Roni Weaver, Charles Hottenstein, Matthew Szapacs, Lee Abberley, LingSing Chen and Xun Wang
Due to high affinity between biotin and avidin, biotinylation is widely used in ligand-binding-assay development for large-molecule bioanalysis. However, biotinylation adds biotin/spacer moiety onto the molecule and may affect the functional activity of the labeled molecule. The current Gyrolab™ immunoassay system requires the capture reagent to be biotinylated as the solid phase comprises a streptavidin-coated-bead column. During method development of a Gyrolab assay for quantification of otelixizumab, we discovered that the response and assay sensitivity was affected by freeze-thaw, which might be related to the type of spacers used between the biotin and the labeled antibody. The hydrophilic Polyethylene Glycol (PEG) spacer enhances water solubility but might be more liable to freeze-thaw compared to antibodies labeled with reagents having only hydrocarbon spacers. The overal.. Read More»
DOI:
10.4172/1948-593X.1000110
Journal of Bioanalysis & Biomedicine received 3099 citations as per Google Scholar report