A modified fast micro method in agarose for isotype, allotype, light chain and idiotype-specific analysis of antibody clonotypes to bacterial virulence antigens.

Nagao AT; Hahn-Zoric M; Wadsworth C; Gustafsson B; Ulanova M; Hanson LA

首页> 外文期刊>Scandinavian journal of clinical and laboratory investigation. >A modified fast micro method in agarose for isotype, allotype, light chain and idiotype-specific analysis of antibody clonotypes to bacterial virulence antigens.

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A modified fast micro method in agarose for isotype, allotype, light chain and idiotype-specific analysis of antibody clonotypes to bacterial virulence antigens.

机译：一种改良的琼脂糖快速微方法，用于针对细菌毒力抗原的抗体克隆型的同种型，同种异型，轻链和独特型特异性分析。

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摘要

A modified fast micro method for spectrotypic/clonotypic analysis of human IgG1-4 antibodies against bacterial virulence antigens of polysaccharide or protein nature is described. Serum samples of as small volumes as 0.5 microliter were isoelectrically focused in micro agarose gels made in plexiglass matrices and blotted using immunoaffinity-mediated capillary blotting onto nitrocellulose membranes previously coated with antigen. The bands of the antigen-specific antibodies were identified with respect to isotypes, light chain types, allotypes or idiotypes by incubating the nitrocellulose membranes with mouse monoclonal anti-human IgG subclass antisera and then with alkaline phosphatase-conjugated rabbit anti-mouse immunoglobulins. The method was applied for characterization of human monoclonals against tetanus toxoid (TT) and for the analysis of variable patterns of clonotypes in IgG subclass-deficient patients. The usefulness of the technique was also demonstrated by comparing the variable specificity and reactivity of different commercial monoclonals against human IgG subclasses. This method is fast, specific, sensitive, uses little material, is simple and reproducible.

机译：描述了一种针对人IgG1-4抗体针对多糖或蛋白质性质的细菌毒力抗原的分型/克隆型分析的改良快速微方法。将体积小至0.5微升的血清样品等电聚焦在有机玻璃基质中制成的微琼脂糖凝胶中，并使用免疫亲和力介导的毛细管印迹法将其印迹到预先涂有抗原的硝酸纤维素膜上。通过将硝酸纤维素膜与小鼠单克隆抗人IgG亚类抗血清，然后与碱性磷酸酶偶联的兔抗小鼠免疫球蛋白一起孵育，就同种型，轻链类型，同种异型或独特型确定了抗原特异性抗体的条带。该方法用于鉴定抗破伤风类毒素（TT）的人类单克隆抗体，并用于分析IgG亚类缺陷患者的克隆型可变模式。通过比较不同商品化单克隆抗体对人IgG亚类的可变特异性和反应性，也证明了该技术的实用性。该方法快速，专一，灵敏，使用的材料少，简单且可重复。

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《Scandinavian journal of clinical and laboratory investigation.》 |1998年第8期|共8页
作者
Nagao AT; Hahn-Zoric M; Wadsworth C; Gustafsson B; Ulanova M; Hanson LA;
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正文语种 eng
中图分类诊断学;
关键词
Antibodies; Bacterial; Antibody Specificity; Antigens; Bacterial; Electrophoresis; Agar Gel; 抗体; 细菌; 抗体特异性; 抗原; 细菌; 电泳; 琼脂凝胶;

机译：Antibodies;Bacterial;Antibody Specificity;Antigens;Bacterial;Electrophoresis;Agar Gel;抗体;细菌;抗体特异性;抗原;细菌;电泳;琼脂凝胶;

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1. A modified fast micro method in agarose for isotype, allotype, light chain and idiotype-specific analysis of antibody clonotypes to bacterial virulence antigens. [J] . Nagao AT, Hahn-Zoric M, Wadsworth C, Scandinavian journal of clinical and laboratory investigation. . 1998,第8期

机译：一种改良的琼脂糖快速微方法，用于针对细菌毒力抗原的抗体克隆型的同种型，同种异型，轻链和独特型特异性分析。
2. In vivo microtubules are copolymers of available beta-tubulin isotypes: localization of each of six vertebrate beta-tubulin isotypes using polyclonal antibodies elicited by synthetic peptide antigens. [J] . M A Lopata, D W Cleveland Journal of cell biology . 1987,第4期

机译：体内微管是可用的β-微管蛋白同种型的共聚物：使用合成肽抗原引发的多克隆抗体定位六个脊椎动物β-微管蛋白同种型中的每一个。
3. Antibody response of rabbit blood lymphocytes in vitro. Kinetics, clone size, and clonotype analysis in response to streptococcal group polysaccharide antigens. [J] . D G Braun, J Quintáns, A L Luzzati, The Journal of Experomental Medicine . 1976,第2期

机译：体外兔血淋巴细胞的抗体反应。响应链球菌多糖抗原的动力学，克隆大小和克隆型分析。
4. Identification of antibody isotypes in biological fluids by means of micro-Raman spectroscopy and chemometric methods [C] . C. Araujo-Andrade, J. L. Pichardo-Molina, G. Barbosa-Sabanero, Conference on Optical Diagnostics and Sensing; 20080121,23; San Jose,CA(US) . 2008

机译：通过微拉曼光谱和化学计量学方法鉴定生物液体中的抗体同种型
5. Comparative analysis of humanized antibodies with small changes in either the heavy chain or light chain of the antibody [D] . Rajendran, Sakthi Harini. 2016

机译：人源化抗体的重链或轻链变化很小的比较分析
6. Evolution of immunoglobulin light chain genes: analysis of Xenopus IgL isotypes and their contribution to antibody diversity. [O] . J Schwager, N Bürckert, M Schwager, 1991

机译：免疫球蛋白轻链基因的进化：非洲爪蟾IgL同种型及其对抗体多样性的贡献的分析。
7. The bacterial identification and antimicrobial susceptibility testing are usually performed manually. Recently, the number of samples has increased and the need for automatification has grown. BioMerieux has investigated Vitek2 for this purpose. HUSLAB has assigned us to compare the identification and antimicrobial results with respect to routine methods and Vitek2. Furthermore, we evaluate test reactions, ESBL (Extended Spectrum Beta-Lactamase) production and the time what it takes to get results with Vitek2. In this study, we examined a total of 183 bacterial isolates including 76 ESBL-strains, 40 nonfermenting rods, 41 fastidious bacteria and 26 fresh clinical isolates. The results of routine methods were gathered from the patient database and following this, compared to the results given by Vitek2. Vitek2 correctly identified more than 96 f the ESBL-strains and fresh clinical isolates. The results of nonfermenting rods and fastidious bacteri a were 70 oncordant with those of the routine methods. Test reaction results of Api20E were 94 nd Api20NE 70 n concordance with Vitek2 results. Approximately 80 f all Vitek2 antimicrobial results and 90 f ESBL test results were correct. Compared to earlier studies, Vitek2 performed the tests faster than the routine methods. These results suggest that Vitek2 is capable of giving rapid and accurate results. However, nonfermenting rods and fastidious bacteria should be studied further. [O] . Järveläinen Marika, Roslund Iira, Silander Maarit 2006

机译：细菌鉴定和抗菌药敏试验通常是手动进行的。最近，样品的数量增加了，并且对自动化的需求也在增长。 BioMerieux为此进行了Vitek2的研究。 HUSLAB已指派我们比较常规方法和Vitek2的鉴定和抗菌结果。此外，我们还评估了测试反应，ESBL（扩展频谱β-内酰胺酶）的产生以及使用Vitek2获得结果所需的时间。在这项研究中，我们检查了总共183个细菌分离株，包括76个ESBL菌株，40个非发酵棒，41个营养细菌和26个新鲜临床分离株。从患者数据库中收集常规方法的结果，然后将其与Vitek2给出的结果进行比较。 Vitek2正确鉴定出超过96种ESBL菌株和新鲜的临床分离株。与常规方法相比，非发酵棒和精制细菌的结果为70。 Api20E的测试反应结果与Vitek2结果一致为94nd Api20NE 70 n。所有Vitek2抗菌结果约80 f和ESBL测试结果90 f正确。与早期的研究相比，Vitek2的测试速度快于常规方法。这些结果表明Vitek2能够给出快速而准确的结果。但是，应进一步研究非发酵棒和营养细菌。

A modified fast micro method in agarose for isotype, allotype, light chain and idiotype-specific analysis of antibody clonotypes to bacterial virulence antigens.

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