Shen H D, Lin W L, Tsai L C, Tam M F, Chua K Y, Chen H L, Hsieh K H, Li C S, Thomas W R
Department of Medical Research, Veterans General Hospital-Taipei, Centre for Immunology, National Yang-Ming University, Taiwan, Republic of China.
Clin Exp Allergy. 1997 Jul;27(7):824-32.
The group 7 mite allergens react with IgE in 50% of sera from allergic patients.
To determine the molecular and antigenic characteristics and heterogeneity of Der f 7 in mite extracts.
Monoclonal antibodies (MoAbs) produced from mice immunized with recombinant Der f 7 were examined for crossreactivity to Der p 7 and then used for immunoblotting of 1 and 2-D gel electrophoresis. Deglycosylation was studied with N-glycosidase-F and N-terminal sequencing by Edman degradation. The epitopes of the monoclonal antibodies were compared by cross-inhibitory immunoassays.
Immunoblotting of D. farinae extracts with all the anti Der f 7 MoAbs showed major reactivities at 31, 30 and 25 kDa. The strongest immunostaining was at 25 kDa which contrasted with Der p 7 where the 31 and 30 kDa bands were strongest. The relative strength of staining however varied between extracts. The 31 and 30 kDa components were glycosylation products of the 25 kDa form which had the N-terminal sequence predicted from cDNA analysis. Two MoAbs stained an 18 kDa band consistent with a degradation product. The 2-D gels showed that different components with pIs from 5.6-6.4. Both species-specific and Der p 7 crossreactive MoAbs were produced and a two-site ELISA assay for detecting group 7 allergen was developed with MoAbs recognizing different epitopes.
Der f 7 has been defined by its natural N-terminal sequence and MoAbs. It apparently exists as different glycosylation and degradation products in mite extracts, the relative abundance of which differs with different preparations. A two-site ELISA to measure the allergen was developed.
7组螨过敏原可与50%过敏患者血清中的IgE发生反应。
确定螨提取物中Der f 7的分子、抗原特性及异质性。
检测用重组Der f 7免疫小鼠产生的单克隆抗体（MoAbs）与Der p 7的交叉反应性，然后用于1D和2D凝胶电泳的免疫印迹分析。用N-糖苷酶-F研究去糖基化，并通过埃德曼降解法进行N端测序。通过交叉抑制免疫分析比较单克隆抗体的表位。
用所有抗Der f 7 MoAbs对粉尘螨提取物进行免疫印迹分析，结果显示在31 kDa、30 kDa和25 kDa处有主要反应性。最强的免疫染色出现在25 kDa处，这与Der p 7不同，Der p 7中31 kDa和30 kDa条带最强。然而，提取物之间染色的相对强度有所不同。31 kDa和30 kDa成分是25 kDa形式的糖基化产物，其N端序列与cDNA分析预测的一致。两种MoAbs对一条18 kDa条带染色，与降解产物一致。2D凝胶显示不同成分的等电点为5.6 - 6.4。产生了种属特异性和与Der p 7交叉反应的MoAbs，并开发了一种用识别不同表位的MoAbs检测7组过敏原的双位点ELISA检测法。
Der f 7已通过其天然N端序列和MoAbs得以明确。它在螨提取物中显然以不同的糖基化和降解产物形式存在，其相对丰度因不同制剂而异。开发了一种用于测量该过敏原的双位点ELISA检测法。
