Priiatkin S A, Nazarov I B, Morozov V I, Rogozkin V A
Vopr Med Khim. 1988 Jul-Aug;34(4):74-8.
Radioimmunoassay (RIA) of human lysozyme was developed using 125I-lysozyme and specific antiserum to the enzyme. Titre of antibodies produced in the antiserum was as high as 1:81,000, affinity constant constituted 1.3.10(9) M/L. Optimum patterns of the RIA were achieved under the following conditions: B0 = 50-55%, T = 25-30.10(3) cpm, BSA = 0.5%, NaCl = 0.85%, charcoal 2.5%. The antibody-antigen reaction was carried out within 30 min, 30 and 15 min at 37 degrees, 20 degrees and 4 degrees, respectively. The RIA enabled to estimate lysozyme with sensitivity of 0.25 ng/ml, while the standard curve was optimal within the range of 0.1-1.0 ng/tube. Intra- and inter-assay coefficients did not exceed 4% and 8%, respectively. Recovery of the standard lysozyme concentration was equal to 96.5 +/- +/- 0.8%. As shown by the RIA intensive physical exercises resulted in an increase of lysozyme content in blood, which appears to occur due to the enzyme secretion from phagocytes. Calculation of the secretion index may be used for evaluation of blood phagocytes functional state.
利用¹²⁵I-溶菌酶和该酶的特异性抗血清建立了人溶菌酶的放射免疫测定法（RIA）。抗血清中产生的抗体效价高达1:81,000，亲和常数为1.3×10⁹ M/L。在以下条件下可实现RIA的最佳模式：B₀ = 50 - 55%，T = 25 - 30×10³ cpm，牛血清白蛋白（BSA） = 0.5%，氯化钠（NaCl） = 0.85%，活性炭2.5%。抗体-抗原反应分别在37℃、20℃和4℃下进行30分钟、30分钟和15分钟。该RIA能够以0.25 ng/ml的灵敏度估计溶菌酶，而标准曲线在0.1 - 1.0 ng/管范围内最佳。批内和批间系数分别不超过4%和8%。标准溶菌酶浓度的回收率为96.5±0.8%。RIA结果显示，高强度体育锻炼导致血液中溶菌酶含量增加，这似乎是由于吞噬细胞分泌该酶所致。分泌指数的计算可用于评估血液吞噬细胞的功能状态。
