Malinowski K, Manski W
J Immunol Methods. 1978;21(1-2):167-77. doi: 10.1016/0022-1759(78)90233-8.
A new method for the quantitative analysis of antigens and antibodies has been based on (1) ultrafiltration of the antigen-antibody precipitates through silver membranes of 0.2 micrometer pore size in a specially designed multisample apparatus, and (2) spectrophotometric determination at 210 nm of the amount of proteins in the antigen-antibody precipitates dissolved in 0.01 N HCl. At this wavelength, the = C = O group of the polypeptide chains constitutes the main chromophoric group. In comparisons with the ninhydrin color reaction, protein determination by low UV spectrophotometry, e.g., at 210 nm, was shown to be about 6 times more sensitive, permitting analysis of samples containing from 1.0 to 35.0 microgram of antigen. The concentration range of protein solutions in 0.01 N HCl which is measured at low UV can be regulated by a factor of 8--10 by changing absorption between 200 and 230 nm. Comparison of the ultrafiltration microtechnique with the standard quantitative precipitin microtechnique involving centrifugation of precipitates was made in 4 different antigen-antibody systems. The new technique was found to be as accurate as the standard technique. It allows completion of analysis within only 5--6 h. The standard precipitin technique, by contrast, requires a 5--7-day reaction period for completion.
（1）在专门设计的多样品装置中，通过孔径为0.2微米的银膜对抗原 - 抗体沉淀物进行超滤；（2）对溶解于0.01N盐酸中的抗原 - 抗体沉淀物中的蛋白质含量在210nm处进行分光光度测定。在此波长下，多肽链的 = C = O基团构成主要发色基团。与茚三酮显色反应相比，通过低紫外分光光度法（例如在210nm处）测定蛋白质的灵敏度约高6倍，可对含1.0至35.0微克抗原的样品进行分析。通过改变200至230nm之间的吸光度，在0.01N盐酸中用低紫外测定的蛋白质溶液浓度范围可调节8 - 10倍。在4种不同的抗原 - 抗体系统中，将超滤微技术与涉及沉淀物离心的标准定量沉淀素微技术进行了比较。发现新技术与标准技术一样准确。它仅需5 - 6小时就能完成分析。相比之下，标准沉淀素技术需要5 - 7天的反应期才能完成。
