Chinese Journal of Dermatology ›› 2012, Vol. 45 ›› Issue (6): 384-387.

• Original articles • Previous Articles     Next Articles

Development of a method to detect anti-BP180NC16A IgG subclasses and the significance of anti-BP180NC16A IgG in bullous pemphigoid

  

  • Received:2011-12-15 Revised:2012-03-16 Online:2012-06-15 Published:2012-05-31
  • Contact: Ya-gang ZUO E-mail:zuoyagang@126.com
  • Supported by:

    ;New-Century Training Programme Foundation for the Talents by Ministry of Education of China

Abstract:

【Abstract】 Objective To develop an assay to quantitatively detect anti-BP180NC16A IgG subclasses and to assess the significance of anti-BP180NC16A IgG in bullous pemphigoid (BP). Methods The Glutathione S-transferase(GST)-BP180NC16A fusion protein was expressed in E.coli system and purified by affinity chromatography. An improved enzyme-linked immunoabsorbent assay (ELISA) was developed and used to detect anti-BP180NC16A IgG subclasses in serum samples from 10 patients with BP, 5 patients with pemphigoid gestationis (PG), 1 patient with linear IgA bullous dermatosis (LIBD) and 2 patients with pemphigus. Results The optimal condition for the ELISA was determined by cross assay as follows: the concentration of GST-BP180NC16A fusion protein for coating, 500 μg/L; the condition for coating, 4 ℃ for 12 hours; the dilution ratio of sera and secondary antibody, 1 ∶ 100 and 1 ∶ 2000 respectively; the condition for incubation, 37 ℃ for 1 hour; the condition for the enzyme-substrate reaction, 37 ℃ for 20 minutes. Of the 10 patients with BP, all were positive for anti-BP180NC16A IgG1, 9 for IgG2, 5 for IgG3, and 9 for IgG4. Anti-BP180NC16A IgG was undetected in any of the serum samples from 2 patients with pemphigus vulgaris or 1 patient with adult LIBD. All the 5 sera from patients with PG were positive for all the anti-BP180NC16A IgG subclasses, which were predominated by IgG1 and IgG3. Conclusion The developed ELISA is a highly specific and reproducible semi-quantitative method for the detection of anti-BP180NC16A IgG subclasses in patients with BP and PG.

Key words: ELISA