Optimizing the Production of gp145, an HIV-1 Envelope Glycoprotein Vaccine Candidate and Its Encapsulation in Guanosine Microparticles

被引:4
作者
Akamine, Pearl [1 ]
Gonzalez-Feliciano, Jose A. [1 ]
Almodovar, Ruth [2 ]
Morell, Gloriner [2 ]
Rivera, Javier [2 ]
Capo-Velez, Coral M. M. [1 ]
Delgado-Velez, Manuel [1 ,3 ]
Prieto-Costas, Luis [4 ]
Madera, Bismark [1 ,3 ]
Eichinger, Daniel [2 ]
Pino, Ignacio [2 ]
Rivera, Jose H. [2 ]
Ortiz-Ubarri, Jose [5 ]
Rivera, Jose M. [4 ]
Baerga-Ortiz, Abel [1 ,6 ]
Lasalde-Dominicci, Jose A. [1 ,3 ,4 ,7 ]
机构
[1] Univ Puerto Rico, Clin Bioreagent Ctr, Mol Sci Res Ctr, San Juan, PR 00926 USA
[2] CDI Labs, Mayaguez, PR 00680 USA
[3] Univ Puerto Rico, Dept Biol, Rio Piedras Campus, San Juan, PR 00931 USA
[4] Univ Puerto Rico, Dept Chem, Rio Piedras Campus, San Juan, PR 00925 USA
[5] Univ Puerto Rico, Dept Comp Sci, Rio Piedras Campus, San Juan, PR 00925 USA
[6] Univ Puerto Rico, Dept Biochem, Med Sci Campus, San Juan, PR 00936 USA
[7] Univ Puerto Rico, Inst Neurobiol, Med Sci Campus, San Juan, PR 00901 USA
基金
美国国家卫生研究院;
关键词
human immunodeficiency virus; envelope protein; gp145; vaccine; glycosylation; encapsulation; microparticles; supramolecular hacky sacks; manufacturing; scale-up; RATIONAL DESIGN; ENV TRIMERS; GLYCOSYLATION; ANTIBODY; PROTEIN; GP120; IMMUNOGENICITY; ANTIGENICITY; EXPRESSION; PROFILES;
D O I
10.3390/vaccines11050975
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
We have developed a pipeline to express, purify, and characterize HIV envelope protein (Env) gp145 from Chinese hamster ovary cells, to accelerate the production of a promising vaccine candidate. First in shake flasks, then in bioreactors, we optimized the growth conditions. By adjusting the pH to 6.8, we increased expression levels to 101 mg/L in a 50 L bioreactor, nearly twice the previously reported titer value. A battery of analytical methods was developed in accordance with current good manufacturing practices to ensure a quality biopharmaceutical. Imaged capillary isoelectric focusing verified proper glycosylation of gp145; dynamic light scattering confirmed the trimeric arrangement; and bio-layer interferometry and circular dichroism analysis demonstrated native-like properties (i.e., antibody binding and secondary structure). MALDI-TOF mass spectrometry was used as a multi-attribute platform for accurate mass determination, glycans analysis, and protein identification. Our robust analysis demonstrates that our gp145 product is very similar to a reference standard and emphasizes the importance of accurate characterization of a highly heterogeneous immunogen for the development of an effective vaccine. Finally, we present a novel guanosine microparticle with gp145 encapsulated and displayed on its surface. The unique properties of our gp145 microparticle make it amenable to use in future preclinical and clinical trials.
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页数:22
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共 52 条
[1]   Rational Design of DNA-Expressed Stabilized Native-Like HIV-1 Envelope Trimers [J].
Aldon, Yoann ;
McKay, Paul F. ;
Allen, Joel ;
Ozorowski, Gabriel ;
Levai, Reka Felfodine ;
Tolazzi, Monica ;
Rogers, Paul ;
He, Linling ;
de Val, Natalia ;
Fabian, Katalin ;
Scarlatti, Gabriella ;
Zhu, Jiang ;
Ward, Andrew B. ;
Crispin, Max ;
Shattock, Robin J. .
CELL REPORTS, 2018, 24 (12) :3324-+
[2]   Cleavage -Independent HIV-1 Trimers From CHO Cell Lines Elicit Robust Autologous Tier 2 Neutralizing Antibodies [J].
Bale, Shridhar ;
Martine, Alexandra ;
Wilson, Richard ;
Behrens, Anna-Janina ;
Le Fourn, Valerie ;
de Val, Natalia ;
Sharma, Shailendra K. ;
Tran, Karen ;
Torres, Jonathan L. ;
Girod, Pierre-Alain ;
Ward, Andrew B. ;
Crispin, Max ;
Wyatt, Richard T. .
FRONTIERS IN IMMUNOLOGY, 2018, 9
[3]   A recombinant human immunodeficiency virus type 1 envelope glycoprotein complex stabilized by an intermolecular disulfide bond between the gp120 and gp41 subunits is an antigenic mimic of the trimeric virion-associated structure [J].
Binley, JM ;
Sanders, RW ;
Clas, B ;
Schuelke, N ;
Master, A ;
Guo, Y ;
Kajumo, F ;
Anselma, DJ ;
Maddon, PJ ;
Olson, WC ;
Moore, JP .
JOURNAL OF VIROLOGY, 2000, 74 (02) :627-643
[4]   Protein-based antigen presentation platforms for nanoparticle vaccines [J].
Brian Nguyen ;
Tolia, Niraj H. .
NPJ VACCINES, 2021, 6 (01)
[5]   Nanoparticle Vaccines for Inducing HIV-1 Neutralizing Antibodies [J].
Brinkkemper, Mitch ;
Sliepen, Kwinten .
VACCINES, 2019, 7 (03)
[6]   cGMP production and analysis of BG505 SOSIP.664, an extensively glycosylated, trimeric HIV-1 envelope glycoprotein vaccine candidate [J].
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. .
BIOTECHNOLOGY AND BIOENGINEERING, 2018, 115 (04) :885-899
[7]   Envelope glycans of immunodeficiency virions are almost entirely oligomannose antigens [J].
Doores, Katie J. ;
Bonomelli, Camille ;
Harvey, David J. ;
Vasiljevic, Snezana ;
Dwek, Raymond A. ;
Burton, Dennis R. ;
Crispin, Max ;
Scanlan, Christopher N. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2010, 107 (31) :13800-13805
[8]   Glycan Masking Focuses Immune Responses to the HIV-1 CD4-Binding Site and Enhances Elicitation of VRC01-Class Precursor Antibodies [J].
Duan, Hongying ;
Chen, Xuejun ;
Boyington, Jeffrey C. ;
Cheng, Cheng ;
Zhang, Yi ;
Jafari, Alexander J. ;
Stephens, Tyler ;
Tsybovsky, Yaroslav ;
Kalyuzhniy, Oleksandr ;
Zhao, Peng ;
Menis, Sergey ;
Nason, Martha C. ;
Normandin, Erica ;
Mukhamedova, Maryam ;
DeKosky, Brandon J. ;
Wells, Lance ;
Schief, William R. ;
Tian, Ming ;
Alt, Frederick W. ;
Kwong, Peter D. ;
Mascola, John R. .
IMMUNITY, 2018, 49 (02) :301-+
[9]   Challenges of glycoprotein analysis by microchip capillary gel electrophoresis [J].
Engel, Nicole ;
Weiss, Victor U. ;
Wenz, Christian ;
Ruefer, Andreas ;
Kratzmeier, Martin ;
Glueck, Susanne ;
Marchetti-Deschmann, Martina ;
Allmaier, Guenter .
ELECTROPHORESIS, 2015, 36 (15) :1754-1758
[10]   An HIV Vaccine Is Essential for Ending the HIV/AIDS Pandemic [J].
Fauci, Anthony S. .
JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION, 2017, 318 (16) :1535-1536