本文選題：蛋白芯片　切入點(diǎn)：β-乳球蛋白　出處：《衛(wèi)生研究》2017年01期

【摘要】：目的優(yōu)化蛋白芯片檢測(cè)β-乳球蛋白(β-Lg)的技術(shù)條件。方法用芯片點(diǎn)樣儀將β-乳球蛋白的一種抗體點(diǎn)樣于三維基片上,用其另一種抗體作為檢測(cè)抗體,以Cy3標(biāo)記的羊抗作為二抗進(jìn)行檢測(cè)。以雙抗夾心法對(duì)β-Lg抗原進(jìn)行檢測(cè)。結(jié)果β-Lg單克隆鼠抗66#被確定為點(diǎn)樣探針,選擇接觸式點(diǎn)樣進(jìn)行芯片點(diǎn)樣,點(diǎn)樣數(shù)在42~92點(diǎn)之間可出現(xiàn)較好的點(diǎn)樣一致性;β-Lg固定探針的濃度選擇0.5 mg/mL,其檢測(cè)抗體的稀釋度為1∶2000;1%無蛋白封閉液被優(yōu)選為封閉劑;β-Lg檢測(cè)下限與生物檢測(cè)限分別為17.54和55.31 ng/mL。根據(jù)β-Lg的S型曲線確定了其線性范圍并建立了對(duì)β-Lg具有最佳決定系數(shù)(R~2=0.9993)的回歸方程與標(biāo)準(zhǔn)曲線。結(jié)論本研究優(yōu)化了牛乳中β-Lg檢測(cè)的蛋白芯片檢測(cè)條件,建立了定量檢測(cè)牛乳β-Lg的蛋白芯片平臺(tái)。
[Abstract]:Objective to optimize the technical conditions for the detection of 尾 -lactoglobulin (尾 -Lg) by using a protein chip. The Cy3 labeled goat antibody was used as the second antibody, and the 尾 -Lg antigen was detected by double antibody sandwich method. The concentration of 尾 -Lg fixed probe was 0.5 mg / mL, the dilution of antibody was 1: 2000 1% and the detection limit of 尾 -Lg was 17.54, while the concentration of 尾 -Lg fixed probe was 17.54, and the concentration of 尾 -Lg fixed probe was 0.5 mg / mL, and the dilution of antibody was 1: 2000 1%, the concentration limit of 尾 -Lg was 17.54, and the concentration of 尾 -Lg was 17.54, and the concentration of 尾 -Lg was 17.54, respectively. And 55.31 ng / mL. According to the S-shape curve of 尾 -Lg, the linear range was determined and the regression equation and standard curve of 尾 -Lg were established. Conclusion the detection conditions of 尾 -Lg in milk by protein chip were optimized. A protein chip platform for quantitative detection of 尾-LG in milk was established.
【作者單位】： 中國疾病預(yù)防控制中心營養(yǎng)與健康所;
【基金】：中國疾病預(yù)防控制中心青年科研基金(No.2015A202)
【分類號(hào)】：R155.5

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本文編號(hào)：1661349