Purification of the integral membrane glycoproteins D of Herpes simplex virus types 1 and 2, produced in the recombinant baculovirus expression system, by ion-exchange high-performance liquid chromatography

Damhof, R.A.; Feijlbrief, M.; Welling‐Wester, Sytske; Welling, Gjalt W.

doi:10.1016/0021-9673(94)80454-0

Cited by 8 publications

(4 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…may be used. Non-denatured protein is usually solubilized in the disruption buffer (Damhof et al, 1994) (possibly with the addition of mild detergents [Welling-Wester et al, 1998]), whereas denatured protein is solubilized with the addition of strong detergents (sodium dodecylsulfate), chaotropes (urea), and possibly reducing agents (2-mercaptoethanol). It is generally preferred to extract non-denatured protein to avoid the complications of developing a suitable renaturation process.…”

Section: Discussionmentioning

confidence: 99%

“…Several chromatographic techniques (such as ion exchange, size exclusion, etc.) have been reported for the purification of VLPs (Damhof et al, 1994;Welling-Wester et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

“…The extraction of recombinant B19 VLPs from the cell paste has proven to be a challenge. Most studies on extraction of baculovirus-infected cells have centered around the use of detergents to facilitate extraction (George et al, 1996;Damhof et al, 1994;Sali et al, 1998;Welling-Wester et al, 1998). In this study we describe the optimized conditions for the extraction of coexpressed VP1 and VP2 from the cell paste.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Enhanced kinetic extraction of parvovirus B19 structural proteins

Sico

White

Tsao

et al. 2002

Biotech & Bioengineering

View full text Add to dashboard Cite

Recombinant structural proteins (VP1 and VP2) of the human parvovirus B19 have been expressed simultaneously using the baculovirus expression system to form virus-like particles (VLPs) that have potential use as vaccines. In this study, we report optimization of extraction conditions to recover these VLPs from cell paste. Under hypotonic conditions with neutral pH these VLPs were poorly extracted (up to 3% extraction). Addition of reducing agents, detergents, salts, and sonication did not improve the extractability. While screening for conditions to improve the extractability of the VLPs, we discovered that a combination of higher pH and elevated processing temperature significantly increased the extraction. Whereas increasing pH alone increased extractability from 3% to 6% (pH increased from 8.0 to 9.5), the effect of elevated temperature was much more substantial. At 50 degrees C, we observed the extraction to be more than fivefold higher than that at room temperature (up to 25% extracted at pH 9.0). The kinetics of extraction at elevated temperatures showed a rapid initial rate of extraction (on the order of minutes) followed by a plateau. In addition, we compared the extraction of VP1 expressed alone. VP1 expressed alone is incapable of forming VLPs. We observed that non-VLP VP1 was easily extractable (up to 60% extracted) under conditions in which the VP1 + VP2 VLPs were not extractable. From these studies we conclude that parvovirus B19 structural proteins expressed to form VLPs have a hindered extractability as compared with non-VLP protein. This hindrance to extraction can be significantly reduced by processing at elevated temperatures and an increased pH, possibly due to the enhanced rates of solubilization and diffusion.

show abstract

Section: Discussionmentioning

confidence: 99%

“…Several chromatographic techniques (such as ion exchange, size exclusion, etc.) have been reported for the purification of VLPs (Damhof et al, 1994;Welling-Wester et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Enhanced kinetic extraction of parvovirus B19 structural proteins

Sico

White

Tsao

et al. 2002

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

“…Peptides that inhibited the binding of mAb A16 to immobilized gD-(Y-l9)-peptide are indicated by +, those that did not inhibit binding are indicated by -. Epitope peptide [gD- (9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)-peptide], LKAhxADPNWRG, where Ahx represents 2-aminohexanoic acid (norleucine). Mimotope peptide, GRFSDALETL.…”

Section: Identification Of Mab-a16-binding Analoguesmentioning

confidence: 99%

Kinetic Analysis of Synthetic Analogues of Linear‐Epitope Peptides of Glycoprotein D of Herpes Simplex Virus Type 1 by Surface Plasmon Resonance

Lasonder¹,

Schellekens²,

Koedijk³

et al. 1996

European Journal of Biochemistry

View full text Add to dashboard Cite

The interaction between mAb A16 and glycoprotein D (gD) of herpes simplex virus type 1 was analyzed by studying the kinetics of binding with a surface-plasmon-resonance biosensor. mAb A1 6 belongs to group VII antibodies, which recognize residues 11 -19 of gD. In a previous study, three critical residues, Aspl3, Argl6 and Phel7, of this epitope were identified by screening a phage display library that contained a random 15-amino-acid insert with the antibody. The contribution to binding of these residues in the motif DXXRF was further analyzed by an amino-acid-replacement study of the epitope gD-(9-19)-peptide and of a gD-(9-19)-peptide mimotope, previously obtained by screening the phage display library. Amino acid residues of the motif were replaced by a neutral amino acid residue, an amino acid residue with opposite charge and a corresponding D-amino acid residue. Kinetic parameters of peptide analogues were determined with a surface plasmon-resonance biosensor. The kinetic parameters of the peptide analogues were compared with the kinetic parameters of the interaction between mAb A16 and the epitope gD-(9-19)-peptide. The minimal size of the gD epitope for mAb A16 was also determined in this study. The kinetic constants of the resulting gD-(ll-l7)-peptide were found to be similar to those of entire gD. The kinetic analysis precisely defined the epitope on gD for mAb A16 to residues 11 -17, identified Argl6 as an essential residue and suggested that Asp13 and Phel7 are mainly involved in stabilization of the secondary structure of the peptide.Keywords: herpes simplex virus 1 ; glycoprotein D; linear epitope ; kinetics ; surface plasmon resonance.Glycoprotein D (gD) of herpes simplex virus (HSV) type 1 is an essential component of the virion envelope for the entry of HSV into mammalian cells [I] and is one of the HSV proteins on which immunological and vaccine studies have focused. Immunization of mice with gD of HSV-1 results in high neutralizing-antibody titers against HSV-1, and these mice are protected against a lethal HSV challenge [ 2 ] . mAbs directed against different regions of gD of HSV-1 have been divided into groups according to their antigenic sites [3], mAbs which recognize amino acid residues 11 -19 in the N-terminus are designated as group VII mAbs [4, 51. mAb A16 is a group VII mAb that binds a synthetic peptide consisting of amino acid residues 9-19 of gD [6, 71. Immunization with synthetic peptides comprising this region of HSV gD protected mice against a lethal challenge with HSV [8, 91. Substitution studies showed that Pro14 and Argl6 are important for binding of mAb A16 [7]. Additional information on residues that interact with mAb A16 was obtained by screening a random peptide library displayed on a filamentous bacteriophage [lo] with the antibody.Peptide sequences obtained by screening such a library resemble or mimic the epitope and are designated as mimotopes. We identified one mimotope. Its smallest synthetic version able to inhibit binding of the gD-(9-19)-peptide to mAb A16 was a ...

show abstract

Mobile Phases and Elution

Wan

2021

Mixed-Mode Chromatography

View full text Add to dashboard Cite

Purification of the integral membrane glycoproteins D of Herpes simplex virus types 1 and 2, produced in the recombinant baculovirus expression system, by ion-exchange high-performance liquid chromatography

Cited by 8 publications

References 32 publications

Enhanced kinetic extraction of parvovirus B19 structural proteins

Enhanced kinetic extraction of parvovirus B19 structural proteins

Kinetic Analysis of Synthetic Analogues of Linear‐Epitope Peptides of Glycoprotein D of Herpes Simplex Virus Type 1 by Surface Plasmon Resonance

Mobile Phases and Elution

Contact Info

Product

Resources

About