التفاصيل البيبلوغرافية
| العنوان: |
cGMP production and analysis of BG505 SOSIP.664, an extensively glycosylated, trimeric HIV‐1 envelope glycoprotein vaccine candidate |
| المؤلفون: |
Dey, Antu K., Cupo, Albert, Ozorowski, Gabriel, Sharma, Vaneet K., Behrens, Anna‐Janina, Go, Eden P., Ketas, Thomas J., Yasmeen, Anila, Klasse, Per J., Sayeed, Eddy, Desaire, Heather, Crispin, Max, Wilson, Ian A., Sanders, Rogier W., Hassell, Thomas, Ward, Andrew B., Moore, John P. |
| المساهمون: |
Bill and Melinda Gates Foundation, National Institutes of Health |
| المصدر: |
Biotechnology and Bioengineering ; volume 115, issue 4, page 885-899 ; ISSN 0006-3592 1097-0290 |
| بيانات النشر: |
Wiley |
| سنة النشر: |
2017 |
| المجموعة: |
Wiley Online Library (Open Access Articles via Crossref) |
| الوصف: |
We describe the properties of BG505 SOSIP.664 HIV‐1 envelope glycoprotein trimers produced under current Good Manufacturing Practice (cGMP) conditions. These proteins are the first of a new generation of native‐like trimers that are the basis for many structure‐guided immunogen development programs aimed at devising how to induce broadly neutralizing antibodies (bNAbs) to HIV‐1 by vaccination. The successful translation of this prototype demonstrates the feasibility of producing similar immunogens on an appropriate scale and of an acceptable quality for Phase I experimental medicine clinical trials. BG505 SOSIP.664 trimers are extensively glycosylated, contain numerous disulfide bonds and require proteolytic cleavage, all properties that pose a substantial challenge to cGMP production. Our strategy involved creating a stable CHO cell line that was adapted to serum‐free culture conditions to produce envelope glycoproteins. The trimers were then purified by chromatographic methods using a 2G12 bNAb affinity column and size‐exclusion chromatography. The chosen procedures allowed any adventitious viruses to be cleared from the final product to the required extent of >12 log 10 . The final cGMP production run yielded 3.52 g (peptidic mass) of fully purified trimers (Drug Substance) from a 200 L bioreactor, a notable yield for such a complex glycoprotein. The purified trimers were fully native‐like as judged by negative‐stain electron microscopy, and were stable over a multi‐month period at room temperature or below and for at least 1 week at 50 ° C. Their antigenicity, disulfide bond patterns, and glycan composition were consistent with trimers produced on a research laboratory scale. The methods reported here should pave the way for the cGMP production of other native‐like Env glycoprotein trimers of various designs and genotypes. |
| نوع الوثيقة: |
article in journal/newspaper |
| اللغة: |
English |
| DOI: |
10.1002/bit.26498 |
| الإتاحة: |
https://doi.org/10.1002/bit.26498Test |
| حقوق: |
http://creativecommons.org/licenses/by/4.0Test/ |
| رقم الانضمام: |
edsbas.F0B21BBA |
| قاعدة البيانات: |
BASE |
