New immunoaffinity beads with zwitterionic moieties to minimize nonspecific protein adsorption and to promote specific target protein capture in serum are prepared. These beads are consisted of poly acrylic acids for enrichment of the binding sites, protein G for controlling antibody orientation, sulfobetaine for reducing the nonspecific protein adsorption in serum, and anti-EpCAM for targeting EpCAM expressed on microvesicles surface. The surfaces of these beads have shown highly hydrophilic characteristics and the surface net charge value was measured close to zero. More than 1.4 time amount of anti-EpCAM was immobilized on these beads compared to that of commercially available beads tethered with protein G, and 50% of distinctive suppression of nonspecific protein adsorption was observed that resulted into doubling the total captured amount of EpCAMs by these beads in serum. These newly developed immunoaffinity beads are suitable for effective isolation of target proteins from serum and are easily applicable to a diagnostic setting.

Novel surface control for efficient isolation of microvesicles in human serum by immunoaffinity capture

244th National Meeting (2012)

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