Gustafsson A
Department of Periodontology, Karolinska Institutet, Stockholm, Sweden.
J Clin Periodontol. 1996 May;23(5):432-6. doi: 10.1111/j.1600-051x.1996.tb00570.x.
The purpose of this study was to assess the recovery of certain proteins from paper strips used for sampling of gingival crevicular fluid (GCF). During a period of 60 min, proteins were eluent in isotonic phosphate-buffered saline, pH 7.4, without detergent. This eluent did not lyse neutrophils applied to the strips, which might otherwise contribute intracellular substances-, e.g., elastase to the GCF samples. The proteins applied (MW 12-725 kD) were satisfactorily recovered (about 90%). However, free neutrophil elastase, unlike elastase complexed with alpha-2-macroglobulin, could not be recovered from the paper strips. Strips made of polyvinylidene difluoride (Durapore) were also tested and 31% of the applied pure elastase was recovered from them. A comparison of measurements made with a Periotron 6000 on Periopaper and Durapore strips, showed that the values obtained with Durapore were too low for accurate measurements of small volumes of fluid. On the whole, when the elution was made in isotonic buffers at neutral pH and without detergents, the recovery of most proteins was satisfactory and reproducible, and independent of MW in the tested range. However, the study also showed poor recovery of uncomplexed elastase.
本研究的目的是评估从用于龈沟液（GCF）采样的纸条中回收某些蛋白质的情况。在60分钟的时间段内，蛋白质在pH 7.4的等渗磷酸盐缓冲盐溶液中洗脱，不使用去污剂。这种洗脱液不会裂解施加在纸条上的中性粒细胞，否则中性粒细胞可能会将细胞内物质（如弹性蛋白酶）释放到GCF样本中。所施加的蛋白质（分子量为12 - 725 kD）回收率令人满意（约90%）。然而，与与α-2-巨球蛋白结合的弹性蛋白酶不同，游离中性粒细胞弹性蛋白酶无法从纸条中回收。还测试了由聚偏二氟乙烯（Durapore）制成的纸条，从其中回收了31%施加的纯弹性蛋白酶。使用Periotron 6000对Periopaper纸条和Durapore纸条进行测量的比较表明，用Durapore纸条获得的值对于准确测量少量液体来说太低了。总体而言，当在中性pH的等渗缓冲液中且不使用去污剂进行洗脱时，大多数蛋白质的回收率令人满意且可重复，并且在所测试的分子量范围内与分子量无关。然而，该研究还表明未结合的弹性蛋白酶回收率较低。
