Purification of baculovirus vectors using heparin affinity chromatography

Abstract

Baculoviruses are commonly used for recombinant protein and vaccine production. Baculoviruses are nonpathogenic to vertebrates, have a large packaging capacity, display broad host and cell type tropism, infect both dividing and nondividing cells, and do not elicit strong immune or allergic responses in vivo. Hence, their use as gene delivery vehicles has become increasingly popular in recent years. Moreover, baculovirus vectors carrying mammalian regulatory elements can efficiently transduce and express transgenes in mammalian cells. Based on the finding that heparan sulfate, which is structurally similar to heparin, is an attachment receptor for baculovirus, we developed a novel scalable baculovirus purification method using heparin-affinity chromatography. Baculovirus supernatants were loaded onto a POROS heparin column, washed to remove unbound materials, and eluted with 1.5 mol/l NaCl, which yielded a recovery of purified baculovirus of 85%. After ultracentrifugation, baculovirus titers increased from 200- to 700-fold with overall yields of 26–29%. We further show that baculovirus particles were infectious, normal in morphology and size, despite high-salt elution and shear forces used during purification and concentration. Our chromatography-based purification method is scalable and, together with ultracentrifugation and/or tangential flow filtration, will be suitable for large-scale manufacturing of baculovirus stocks for protein and vaccine production and in gene therapy applications.

Novel coronavirus-like particles targeting cells lining the respiratory tract

“…The quality of preparation was high, but unfortunately BVs co-purified with VLPs (data not shown). This observation is in agreement with known BVs affinity for heparin columns [ 66 ]. However, this problem was solved by further fractionation using monolith ion exchange column.…”

Polymer brush‐grafted monolithic macroporous polyHIPEs obtained by surface‐initiated ARGET ATRP and heparinized for Enterovirus 71 purification

“…Heparin is a sulfate glycosaminoglycan composed of glucosamine and uronic acid residues alternately, and the structure of heparin was shown in Scheme 1. Heparin has been immobilized onto different matrices for biological application especially in bioseparation 42–45 . In this study, amination of polyHIPE‐ g ‐PGMA was implemented by ring opening reaction with ethylenediamine, and then heparinized by EDC and NHS induced coupling reaction.…”

“…All the results indicated that the polyHIPE‐ g ‐PGMA4‐Hep presented a specific capture of EV71 and verified that it was heparin that played a decisive role in EV71 purification. The specific capture of EV71 was originated from the receptor‐ligand interaction between heparin and EV71 42,53 …”

“…Heparin has been immobilized onto different matrices for biological application especially in bioseparation. [42][43][44][45] In this study, amination of polyHIPE-g-PGMA was implemented by ring opening reaction with ethylenediamine, and then heparinized by EDC and NHS induced coupling reaction. This modification process was confirmed by FTIR and XPS.…”

“…The specific capture of EV71 was originated from the receptor-ligand interaction between heparin and EV71. 42,53…”