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IgA分子大小对其采用各种技术进行免疫测定的影响。II. 固相放射免疫测定

Influence of molecular size of IgA on its immunoassay by various techniques. II. Solid-phase radioimmunoassays.

作者信息

Delacroix D L, Dehennin J P, Vaerman J P

出版信息

J Immunol Methods. 1982;48(3):327-37. doi: 10.1016/0022-1759(82)90333-7.

Abstract

Homogeneous, polymeric (P), monoclonal (MC) and polyclonal (PC) samples of purified IgA, as well as of secretory IgA (sIgA), were compared to their corresponding monomeric (M) forms with regard to their respective behaviour in both a solid-phase double antibody (AB) immunoradiometric assay (IRMA) and a solid-phase competition radioimmunoassay (CRIA). On an identical weight basis, both assays underestimated the P forms. Correction factors (CF), i.e., optical density (OD) versus radiometric concentration ratios, were calculated for both methods for all P forms, using M as standards. IRMA CF were respectively 1.50 for dimer, 1.98 for secretory IgA and 2.40 for tetramers, very similar to those obtained in radial immunodiffusion (RID). With optimally coated beads, these CF were stable along a wide range of concentrations. In contrast, CRIA-CF were 3-4 times larger and displayed much variation along their standard range of concentrations, preventing the use of a constant CF along the standard curve. Our present IRMA, with its CF, should be of value for a more accurate quantitation of small amounts of IgA of various known sizes.

摘要

将纯化的IgA以及分泌型IgA(sIgA)的均一、聚合(P)、单克隆(MC)和多克隆(PC)样本与其相应的单体(M)形式在固相双抗体(AB)免疫放射分析(IRMA)和固相竞争放射免疫分析(CRIA)中的各自行为进行了比较。在相同重量基础上,两种分析方法均低估了P形式。以M为标准品,为所有P形式计算了两种方法的校正因子(CF),即光密度(OD)与放射浓度之比。IRMA的CF对于二聚体分别为1.50,对于分泌型IgA为1.98,对于四聚体为2.40,与在放射免疫扩散(RID)中获得的结果非常相似。使用最佳包被的磁珠,这些CF在很宽的浓度范围内是稳定的。相比之下,CRIA - CF大3 - 4倍,并且在其标准浓度范围内显示出很大变化,这使得无法在标准曲线上使用恒定的CF。我们目前的IRMA及其CF对于更准确地定量各种已知大小的少量IgA应该是有价值的。

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