Production of Leptospiral LipL32 Antigen in Pichia pastoris and Its Use in an Enzyme-Linked Immunosorbent Assay

نویسندگان

  • Leonardo Garcia Monte
  • Fernanda Munhoz Dos Anjos Leal
  • Daiane Drawanz Hartwig
  • Sílvio Arruda Vasconcellos
  • Bibiana Brihuega
  • Odir Antonio Dellagostin
  • Cláudia Pinho Hartleben
چکیده

The production of recombinant LipL32 protein using Escherichia coli has been used extensively for the development of vaccines and diagnostic tests for leptospirosis. However, E. coli has demonstrated limitations, including low yield and lack of post-translational modifications. In this study, rLipL32 was produced in eukaryotic expression system (Pichia pastoris) and evaluated the antigen by enzyme-linked immunosorbent assay (ELISA). The yield obtained from the culture supernatant reached 270 mg/L and ELISA showed an accuracy of 95.34%. In summary, the production of rLipL32 using P. pastoris did not impair the antigenic characteristics of this antigen and ensured its use for detecting the leptospiral antibodies in swine sera.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Heterologous Expression of Bovine Prochymosin in Pichia pastoris GS115

Objectives: In present research we evaluate the expression of this critical enzyme in a eukaryotic system for future use in cheese industry. Materials and Methods: We have cloned bovine prochymosin gene in methylotrophic yeast, P. pastoris, using pPIC9K as an expression vector. The recombinant plasmid was transformed into the host by electroporation, and it was expre...

متن کامل

Evaluation of the recombinant LipL32 in enzyme-linked immunosorbent assay for the serodiagnosis of bovine leptospirosis.

The recombinant leptospiral protein LipL32 was evaluated for use in the diagnosis of bovine leptospirosis by enzyme-linked immunosorbent assay (rLipL32 IgG ELISA). The microscopic agglutination test (MAT) of 150 serum samples from cattle suspected of leptospirosis showed that 125 (83.3%) samples had positive reciprocal agglutination titres, which ranged from 100 to 1600. The highest titres were...

متن کامل

Immunodiagnosis of Haemonchus contortus infection in sheep by indirect enzyme linked immunosorbent assay (ELISA)

Indirect plate enzyme-linked immunosorbent assay was standardized and evaluated for its effectivenessin immunodiagnosis of haemonchosis in experimental and clinical cases in sheep by using somatic wholeadult antigen of H. contortus. Plate ELISA was standardized using 5 μg/well antigen concentration with1:100 and 1:1000 of sera and conjugate dilution. Indirect plate ELISA was able to demonstrate...

متن کامل

Standardization of an Enzyme-Linked Immunosorbent Assay for Detection of Infectious Bronchitis Virus Antibody.

An indirect enzyme–linked immunosorbent assay (ELISA) was developed for screening of antibody to avian infectious bronchitis virus (IBV). Antigen was prepared from whole-purified IBV Massachusetts serotype (BR 801 strain). Optimum dilution with minimum background for antigen concentration, rabbit anti-chicken conjugate and sera in developed ELISA was determined 0.1μg/ml, 1:3000 and 1:100, respe...

متن کامل

Evaluation of Enzyme Linked Immunosorbent Assay, Utilizing Native Antigen B for Serodiagnosis of Human Hydatidosis

Background: Hydatidosis is one of the cosmopolitan parasitic zoonoses caused by the larval stage of Echinococcus granulosus. Diagnosis of hydatidosis is still an unresolved problem. Serological tests using crude antigens for diagnosis of E. granulosus are sensi-tive, however their specificity are not satisfactory. Therefore, WHO recommended spe-cific serological methods using specific antigens,...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره   شماره 

صفحات  -

تاریخ انتشار 2014