本文選題：橢偏光蛋白芯片技術　切入點：光學蛋白質芯片　出處：《山東醫(yī)藥》2016年47期

【摘要】：目的優(yōu)化橢偏光蛋白芯片技術定量檢測血清載脂蛋白B(ApoB)的條件,并觀察最佳條件下橢偏光蛋白芯片技術定量檢測血清ApoB的準確性。方法用橢偏光蛋白芯片技術檢測含有0.2、0.3、0.4、0.5、0.6 mg/m L的載脂蛋白B抗體(anti ApoB)和200μg/m L ApoB標準品的溶液,測得灰度值最高者選為最佳anti-ApoB濃度。用橢偏光蛋白芯片技術檢測5、10、20、50、100、200μg/m L ApoB標準品溶液,anti-ApoB濃度為0.3 mg/m L,結果以灰度值表示。然后在上述標準品溶液中加入蛋白G使其終濃度為0.1 mg/m L,再檢測一次灰度值。比較兩次檢測的灰度值,確定是否應該加入0.1 mg/m L蛋白G。最佳條件下橢偏光蛋白芯片技術檢測0、5、10、20、50、100、200μg/m L ApoB標準品溶液,計算上述ApoB標準品溶液灰度值與不含ApoB的空白探針溶液(含anti ApoB和蛋白G)灰度值的差值。對上述ApoB標準品溶液濃度和灰度值差值的相關性進行分析,以ApoB標準品溶液濃度為橫坐標,灰度值差值為縱坐標描畫標準曲線并建立方程。最佳條件下用橢偏光蛋白芯片技術檢測16份40倍稀釋的已知ApoB濃度(采用免疫比濁法測定)血清的灰度值兩次。用上述方程計算16份血清中ApoB濃度,并與其實際ApoB濃度比較。結果最佳anti-ApoB濃度為0.3 mg/m L。與不應用蛋白G者相比,應用0.1 mg/m L蛋白G的橢偏光蛋白芯片技術測得樣品的灰度值高(P均0.05)。以ApoB濃度為橫坐標x、灰度差值為縱坐標y描畫標準曲線,Maple9.5軟件得出的回歸方程為:y=-23.116Ln(1-x/60),R2=0.936 4。16份血清ApoB濃度和第一次、第二次用橢偏光蛋白芯片技術檢測測得血清ApoB濃度相比P均0.05。結論橢偏光蛋白芯片技術檢測血清ApoB的最佳條件為anti-ApoB濃度0.3 mg/m L,蛋白G濃度0.1 mg/m L。此條件下橢偏光蛋白芯片技術檢測血清ApoB結果準確。
[Abstract]:Objective to optimize the conditions for quantitative detection of serum apolipoprotein (ApoB) by ellipsometry protein chip technique and to observe the accuracy of ellipsometry protein chip technology for quantitative detection of serum ApoB.Methods the solution of Apolipoprotein B antibody anti ApoB and 200 渭 g / mL ApoB standard was detected by ellipsometry protein chip technique. The best anti-ApoB concentration was selected as the highest grayscale value of Apolipoprotein B antibody (ApoB) and 200 渭 g / mL ApoB.The concentration of anti-ApoB in ApoB standard solution was 0.3 mg/m / L using ellipsometry protein chip technique. The results were expressed as gray scale.The final concentration of protein G was 0. 1 mg/m / L, and the gray value was detected again.The gray values of the two tests were compared to determine whether 0.1 mg/m L protein G should be added.Under the optimum conditions, the standard solution of ApoB was detected by ellipsometry protein chip technique. The difference between the gray value of the ApoB standard solution and that of the blank probe solution (including anti ApoB and protein G) without ApoB was calculated.The correlation between the concentration of the ApoB standard solution and the gray value difference was analyzed. The standard curve was drawn and the equation was established with the concentration of the ApoB standard solution as the horizontal coordinate and the gray value difference as the vertical coordinate.Under the optimum conditions, the gray values of 16 serum samples with 40 times dilution of known ApoB concentration (determined by immune turbidimetry) were determined twice by ellipsometry protein chip technique.The ApoB concentration in 16 serum samples was calculated by the above equation and compared with the actual ApoB concentration.Results the optimum concentration of anti-ApoB was 0.3 mg/m L.Compared with those without protein G, the grayscale values of 0. 1 mg/m L protein G were measured by ellipsometry protein chip technique.浠poB嫻撳害涓烘í鍧愭爣x,鐏板害宸，

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