مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

video

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

sound

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Persian Version

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

View:

85
مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Download:

96
مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Cites:

Information Journal Paper

Title

Optimization of Expression and Purification of Recombinant Mouse plac1

Pages

  61-69

Abstract

 Background: Placenta-specific 1 (PLAC1) is one of the recently-discovered Cancer-Testis-Placenta (CTP) antigen with restricted normal tissue and ectopic Expression in a wide range of cancer cells from different histological origins. The production of recombinant human PLAC1 has already been optimized; however, no study has been reported so far on the production and Purification of Mouse plac1. In this study, Mouse plac1 Expression and Purification was optimized in a prokaryotic system and the effects of the generated proteins on inducing humoral responses in mice were investigated. Methods: A fusion protein containing full extracellular domain of Mouse plac1, immunostimulatory peptides, tetanus toxin P2P30 and PADRE and KDEL3 signal (main plac1), and the same fragment without immunostimulatory peptides (control plac1) was produced. To optimize production and Purification steps, different parameters including bacterial strain, cultivation temperature, cultivation time, IPTG concentration, culture medium, and also different buffers for Purification of the Recombinant proteins were tested. After confirming the identity of recombinant plac1 proteins with Western Blotting (WB) and ELISA assays, these proteins were subcutaneously injected in mice with Freund's adjuvant and the anti-plac1 antibody response was detected by ELISA. Results: The optimal Expression level of main and control plac1 was obtained in BL21 (DE3) and TB culture medium in the presence of 0. 25 mM IPTG after 24 hr of induction at 15 C. The buffer containing 2% sarkosyl produced higher yield and purity. Our results showed specific reactivity of anti-human recombinant plac1 polyclonal antibody with both main and control plac1 Recombinant proteins in WB and ELISA analysis. Both proteins induced humoral responses in mice; however, anti-plac1 antibody titer was significantly higher in sera of mice immunized with main compared to control plac1. Conclusion: In this study, an optimized protocol for production and Purification of Mouse plac1 was reported and it was shown that insertion of immunostimulatory peptides in gene construct could efficiently enhance humoral immune responses against Mouse plac1, which could potentially augment cellular immune responses against plac1 leading to more effective anti-cancer responses.

Multimedia

  • No record.

    • Cites

  • No record.

    • References

  • No record.

    • Cite

    APA: Copy

    Rahdan, Shaghayegh, Razavi, Seyed Alireza, NAZARI, MAHBOOBEH, Shojaeian, Sorour, SHOKRI, FAZEL, AMIRI, MOHAMMAD MEHDI, RAMEZANI, AMIN, & ZARNANI, AMIR HASSAN. (2022). Optimization of Expression and Purification of Recombinant Mouse plac1. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), 14(1), 61-69. SID. https://sid.ir/paper/981049/en

    Vancouver: Copy

    Rahdan Shaghayegh, Razavi Seyed Alireza, NAZARI MAHBOOBEH, Shojaeian Sorour, SHOKRI FAZEL, AMIRI MOHAMMAD MEHDI, RAMEZANI AMIN, ZARNANI AMIR HASSAN. Optimization of Expression and Purification of Recombinant Mouse plac1. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB)[Internet]. 2022;14(1):61-69. Available from: https://sid.ir/paper/981049/en

    IEEE: Copy

    Shaghayegh Rahdan, Seyed Alireza Razavi, MAHBOOBEH NAZARI, Sorour Shojaeian, FAZEL SHOKRI, MOHAMMAD MEHDI AMIRI, AMIN RAMEZANI, and AMIR HASSAN ZARNANI, “Optimization of Expression and Purification of Recombinant Mouse plac1,” AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), vol. 14, no. 1, pp. 61–69, 2022, [Online]. Available: https://sid.ir/paper/981049/en

    Related Journal Papers

  • No record.

    • Related Seminar Papers

  • No record.

    • Related Plans

  • No record.

    • Recommended Workshops






    Move to top
    telegram sharing button
    whatsapp sharing button
    linkedin sharing button
    twitter sharing button
    email sharing button
    email sharing button
    email sharing button
    sharethis sharing button