Centre for Contact Lens Research, School of Optometry, University of Waterloo, Waterloo, ON, Canada.
Eye Contact Lens. 2009 Mar;35(2):76-80. doi: 10.1097/ICL.0b013e318199aff6.
Extracting proteins from conventional hydrogel (CH) and silicone hydrogel (SH) contact lens materials using a mixture of trifluoroacetic acid/acetonitrile (TFA/ACN) is a well-established procedure for quantifying individual and total protein deposited on contact lenses. The purpose of this study was to determine the efficacy of TFA/ACN in extracting albumin from SH and a CH group IV lens material using an in vitro model.
One CH group IV lens material (etafilcon A) and five different SH lens materials (lotrafilcon A, lotrafilcon B, balafilcon A, galyfilcon A, and senofilcon A) were incubated in both simple albumin solution and a complex artificial tear protein solution containing 125I-labeled albumin. All the lens materials were incubated for 14 days at 37 degrees C with constant rotations. Following the incubation period, radioactive counts were determined and the lenses were placed in an appropriate volume of the extraction solvent. After the specified time, the lenses were removed and radioactive counts were determined again to calculate the amount of albumin remaining on the lenses post-extraction.
Extraction efficiencies for albumin from the artificial tear protein solution were 97.2% +/- 2 for etafilcon A, 77.3% +/- 6.2 for lotrafilcon A, 73.5% +/- 5.6 for lotrafilcon B, 81.5% +/- 5.8 for balafilcon, 91.2% +/- 3.4 for galyfilcon A, and 89.2% +/- 3.4 for senofilcon A. Results were similar for the albumin extracted after incubating in the simple albumin solution.
Although TFA/ACN is efficient at extracting albumin deposited on etafilcon lenses, it does not extract all the albumin that is deposited on SH lenses and alternative extraction procedures should be sought.
用三氟乙酸/乙腈(TFA/ACN)混合物从传统水凝胶(CH)和硅水凝胶(SH)隐形眼镜材料中提取蛋白质,是一种定量分析隐形眼镜上沉积的个体和总蛋白质的成熟方法。本研究的目的是使用体外模型确定 TFA/ACN 从 SH 和 CH 第四组镜片材料中提取白蛋白的效果。
将一种 CH 第四组镜片材料(etafilcon A)和五种不同的 SH 镜片材料(lotrafilcon A、lotrafilcon B、balafilcon A、galyfilcon A 和 senofilcon A)分别浸泡在简单的白蛋白溶液和含有 125I 标记白蛋白的复杂人工泪液蛋白溶液中。所有镜片材料均在 37°C 下以恒定的转速孵育 14 天。孵育期结束后,测定放射性计数,并将镜片置于适当体积的提取溶剂中。指定时间后,取出镜片并再次测定放射性计数,以计算提取后镜片上残留的白蛋白量。
从人工泪液蛋白溶液中提取白蛋白的提取效率为 etafilcon A 为 97.2%±2,lotrafilcon A 为 77.3%±6.2,lotrafilcon B 为 73.5%±5.6,balafilcon 为 81.5%±5.8,gallyfilcon A 为 91.2%±3.4,senofilcon A 为 89.2%±3.4。在简单白蛋白溶液中孵育后提取的白蛋白结果相似。
尽管 TFA/ACN 能够有效地提取沉积在 etafilcon 镜片上的白蛋白,但它并不能提取沉积在 SH 镜片上的所有白蛋白,因此应寻求其他提取方法。