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Construction of chimeric proteins between protein G and fluorescence-enhanced green fluorescent protein, and their application to immunoassays

, : Construction of chimeric proteins between protein G and fluorescence-enhanced green fluorescent protein, and their application to immunoassays. Journal of Fermentation & Bioengineering 86(5): 440-445

Chimeric proteins consisting of an IgG-binding domain of protein G (PG) and a fluorescence-enhanced green fluorescent protein variant (EGFP) were constructed to evaluate their utility in fluoroimmunoassays (FIAs). Four chimeric proteins (PG-EGFPs) with an amino acid residue of Ala, Pro, Ser or Thr as the linker between the two domains were constructed and expressed in E. coli. They were efficiently expressed and could be purified to homogeneity with one-step IgG-affinity chromatography. The proteins retained similar fluorescence spectra to those of EGFP, and the binding constants, as evaluated using BIAcore, revealed their high affinity for rabbit IgG, goat IgG, sheep IgG and mouse IgG, PG-EGFP with Ala, Pro, Ser as the linker showed a high affinity for rabbit IgG while that with Thr had a 3-fold lower binding constant. When PG-EGFPs were applied to sandwich FIAs, a minimum detection limit of 0.1 mug/ml of BSA using anti-BSA antibody, or 1 pg/ml of rabbit IgG was achieved using microplate format. This detection sensitivity was comparable to that obtained using FITC with similar sensitivity. These data show the applicability of these easily prepared PG-EGFPs to various FIAs for sensitive detection of antigens.


DOI: 10.1016/s0922-338x(98)80148-7

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Milewski, J.B.; Krzeski, T.; Willak, J., 1987: Effect of treatment with mineral water from the Dabrówka spring at the Szczawno-Zdròj health resort on uric acid levels of the blood and urine. Wiadomosci Lekarskie 40(13): 875-878

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Alvarez, R.E., 2004: Lung field segmenting in dual-energy subtraction chest X-ray images. The purpose of this study was to develop and test a method to delineate lung field boundaries in dual-energy chest x-ray images. The segmenting method uses soft-tissue images and spatial frequency-dependent, background-subtracted images. Large-sca...

Kerr, T.; Williams, M.1; Tritz, G.1; Washam, C., 1983: Purification and identification of a nicotinic acid-replacing substance from vitamin-free acid hydrolyzed casein Dairy product. Journal of dairy science 66(4): 750-755

Bracken, M.B., 2016: Case-control studies require appropriate population controls: an example of error in the SSRI birth defect literature. European Journal of Epidemiology 30(11): 1217-1218

Drake Lon D., 1972: Mechanisms of Clast Attrition in Basal Till. Abrasion, crushing, east-central New Hampshire.

Milne, E.N.; Bass, H., 1969: Relationship between specific dynamic pulmonary compliance and diaphragmatic excursion. Radiology 92(3): 615-615

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