High-Throughput Baculovirus Expression in Insect Cells

2011 ◽  
pp. 609-627 ◽  
Author(s):  
Richard B. Hitchman ◽  
Robert D. Possee ◽  
Linda A. King
Keyword(s):  
High Throughput ◽  
Insect Cells ◽  
Baculovirus Expression

scholarly journals Production, processing and partial purification of functional G protein βγ subunits in baculovirus-infected insect cells

1992 ◽  
Vol 286 (3) ◽  
pp. 677-680 ◽  
Author(s):  
J D Robishaw ◽  
V K Kalman ◽  
K L Proulx
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Insect Cells ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Partial Purification ◽  
Baculovirus Expression ◽  
Gamma Subunits ◽  
Infected Insect ◽  
Beta 2

As a result of the inability to resolve the heterogeneous mixture of G protein beta gamma subunits present in tissues, it has not been possible to compare different beta gamma subunits of the G proteins in terms of their proposed roles in receptor-effector coupling. This study was undertaken to establish the utility of the baculovirus expression system in producing homogeneous beta gamma subunits of defined composition for the comparative analysis of these subunits in reconstitution systems. In this study we report the expression, and appropriate post-translational processing, of recombinant beta 2, gamma 2 and gamma 3 subunits. In addition, we show that the recombinant beta gamma subunits can be readily purified, and can functionally interact with the alpha subunits of the G proteins.

scholarly journals Construction of gateway-compatible baculovirus expression vectors for high-throughput protein expression and in vivo microcrystal screening

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yanyang Tang ◽  
Justin Saul ◽  
Nirupa Nagaratnam ◽  
Jose M. Martin-Garcia ◽  
Petra Fromme ◽  
...  
Keyword(s):  
Protein Expression ◽  
High Throughput ◽  
Expression Vectors ◽  
Baculovirus Expression

scholarly journals Partial characterization of natural and recombinant human soluble CD23

1992 ◽  
Vol 286 (3) ◽  
pp. 819-824 ◽  
Author(s):  
K Rose ◽  
G Turcatti ◽  
P Graber ◽  
S Pochon ◽  
P O Regamey ◽  
...  
Keyword(s):  
Insect Cells ◽  
Peptide Mapping ◽  
Sequence Similarity ◽  
Expression System ◽  
Protein A ◽  
Baculovirus Expression System ◽  
Partial Characterization ◽  
Disulphide Bonds ◽  
Baculovirus Expression

The purification to homogeneity of an active soluble 25 kDa fragment of CD23, produced in insect cells using the baculovirus expression system, is described. Peptide mapping and analysis by Edman degradation and mass spectrometry permitted partial characterization of the protein. A total of 165 out of 172 residues, including N-terminal and C-terminal regions, were mapped. The positions of the two disulphide bonds in the IgE-binding region were also determined: residue 110 is joined to residue 124, and residue 42 to residue 133. Natural CD23 25 kDa fragment was also analysed and found to possess the same disulphide bond arrangement. These results extend the previously noted sequence similarity with lectins to elements of secondary structure.

Strong buffering capacity of insect cells. Implications for the baculovirus expression system

1995 ◽  
Vol 17 (1) ◽  
pp. 21-26 ◽  
Author(s):  
Miguel Medina ◽  
Abelardo L�pez-Rivas ◽  
Douwe Zuidema ◽  
Graham J. Belsham ◽  
Esteban Domingo ◽  
...  
Keyword(s):  
Insect Cells ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Buffering Capacity ◽  
Baculovirus Expression

scholarly journals Unique features of a recombinant haemagglutinin influenza vaccine that influence vaccine performance

npj Vaccines ◽  
2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Arun B. Arunachalam ◽  
Penny Post ◽  
Deborah Rudin
Keyword(s):  
Influenza Vaccine ◽  
Adverse Reactions ◽  
Insect Cells ◽  
Age Groups ◽  
Structural Features ◽  
Vector System ◽  
Baculovirus Expression Vector System ◽  
Baculovirus Expression ◽  
Baculovirus Expression Vector ◽  
Protective Antibodies

AbstractThe influenza vaccine field has been constantly evolving to improve the speed, scalability, and flexibility of manufacturing, and to improve the breadth and longevity of the protective immune response across age groups, giving rise to an array of next generation vaccines in development. Among these, the recombinant influenza vaccine tetravalent (RIV4), using a baculovirus expression vector system to express recombinant haemagglutinin (rHA) in insect cells, is the only one to have reached the market and has been studied extensively. We describe how the unique structural features of rHA in RIV4 improve protective immune responses compared to conventional influenza vaccines made from propagated influenza virus. In addition to the sequence integrity, characteristic of recombinant proteins, unique post-translational processing of the rHA in insect cells instills favourable tertiary and quaternary structural features. The absence of protease-driven cleavage and addition of simple N-linked glycans help to preserve and expose certain conserved epitopes on HA molecules, which are likely responsible for the high levels of broadly cross-reactive and protective antibodies with rare specificities observed with RIV4. Furthermore, the presence of uniform compact HA oligomers and absence of egg proteins, viral RNA or process impurities, typically found in conventional vaccines, are expected to eliminate potential adverse reactions to these components in susceptible individuals with the use of RIV4. These distinct structural features and purity of the recombinant HA vaccine thus provide a number of benefits in vaccine performance which can be extended to other viral targets, such as for COVID-19.

Sign in / Sign up

Export Citation Format

Share Document