Expression and characterization of monomeric variable lymphocyte receptor B specific to the glycoprotein of viral hemorrhagic septicemia virus (VHSV)
- Lee, Jung Seok; Kim, Jaesung; Im, Se Pyeong; Kim, Si Won; Jung, Jae Wook; Lazarte, Jassy Mary S.; Lee, Jeong-Ho; Thompson, Kim D.; Jung, Tae Sung
- Issue Date
- ELSEVIER SCIENCE BV
- Variable lymphocyte receptor B (VLRB); Hagfish; Viral hemorrhagic septicemia virus (VHSV); Heterogeneous expression; Pichia pastoris
- JOURNAL OF IMMUNOLOGICAL METHODS, v.462, pp.48 - 53
- Journal Title
- JOURNAL OF IMMUNOLOGICAL METHODS
- Start Page
- End Page
- Monomeric variable lymphocyte receptor B (VLRB) is one of the smallest binding scaffold (20-25 kDa) from jawless vertebrates, hagfish and lamprey. This relatively new class of binding scaffold has various advantages: i) it has a single peptide composition, amenable to molecular engineering for enhancing its stability and affinity; ii) it has a small size, contributing better tissue penetration and easier production using microorganism expression system. Monomeric arVLRB142, which can specifically bind to the glycoprotein of viral hemorrhagic septicemia virus (VHSV), was expressed in Pichia pastoris. High quantity recombinant monomeric arVLRB142 (rVLR142(mono)) was purified from 100 ml of culture with a resulting yield of 2.6 +/- 1.3 mg of target protein. Functional studies revealed that the purified rVLR142(mono) can specifically recognize low levels of the target antigen (recombinant glycoprotein) (i.e. as low as 0.1 nM), but also the native glycoprotein of VHSV. The expressed rVLR142(mono) exhibited high levels of stability and it retained it binding capacity over broad temperature (4 degrees C similar to 60 degrees C) and pH ranges (pH 1.5-12.5). We developed an effective expression system for mass production of monomeric VLRB based on P. pastoris. The recombinant protein that was obtained offers promising binding avidity and biophysical stability and its potential use in various biotechnological applications.
- Files in This Item
- There are no files associated with this item.
- Appears in
- 수의과대학 > Department of Veterinary Medicine > Journal Articles
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.