Abstract
The variable lymphocyte receptors (VLRs) consist of leucine rich repeats (LRRs) and comprise the humoral antibodies produced by lampreys and hagfishes. The diversity of the molecules is generated by stepwise genomic rearrangements of LRR cassettes dispersed throughout the VLRB locus. Previously, target-specific monovalent VLRB antibodies were isolated from sea lamprey larvae after immunization with model antigens. Further, the cloned VLR cDNAs from activated lamprey leukocytes were transfected into human cell lines or yeast to select best binders. Here, we expand on the overall utility of the VLRB technology by introducing it into a filamentous phage display system. We first tested the efficacy of isolating phage into which known VLRB molecules were cloned after a series of dilutions. These experiments showed that targeted VLRB clones could easily be recovered even after extensive dilutions (1 to 109). We further utilized the system to isolate target-specific “lampribodies” from phage display libraries from immunized animals and observed an amplification of binders with relative high affinities by competitive binding. The lampribodies can be individually purified and ostensibly utilized for applications for which conventional monoclonal antibodies are employed.
Highlights
Agnathans diverged from a common vertebrate lineage in the Cambrian period some 500 million years ago [1], with lampreys and hagfish being the only surviving representatives of this group.A unique adaptive immune response mediated by single solenoid-shaped molecules synthesized from highly polymorphic leucine rich repeats (LRR) has been described in lamprey [2] and hagfish [3].These molecules are expressed in cells resembling B-lymphocytes and are called variable lymphocyte receptor B (VLRBs)
In order to examine target specific binding of the VLRB molecules expressed in a phage display system, we used the previously published variable regions of anti-lysozyme VLRB molecules, VLRB.Hen Egg Lysozyme (HEL).1, VLRB.HEL21, and VLRB.HEL2D [9]
PolyA RNA was converted to cDNA which was used as a template for preparation of VLRB amplicon library by PCR with PD-VLRB.F and PD-VLRB.R primers. (C) Agarose gel (1.5%)
Summary
A unique adaptive immune response mediated by single solenoid-shaped molecules synthesized from highly polymorphic leucine rich repeats (LRR) has been described in lamprey [2] and hagfish [3]. These molecules are expressed in cells resembling B-lymphocytes and are called variable lymphocyte receptor B (VLRBs). The system has the theoretical potential of generating a repertoire >1014 different antigen receptors, which is comparable to what is seen in the immunoglobulin (Ig)-based adaptive immune system in the jawed vertebrates [3,6]. VLRB monoclonal antibodies, hitherto referred to as “lampribodies”, have been produced using soluble protein, particulate antigens, and whole cells as immunogens, e.g., BclA protein of Bacillus anthracis exosporium [7], H-trisaccharide of human O-type erythrocyte [8], hen egg lysozyme [9], avian influenza virus hemagglutinin [10,11], CD38 plasma cells [11], plant pathogen protein HopM1 [12], and murine
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
