Generation of Lamprey Monoclonal Antibodies (Lampribodies) Using the Phage Display System

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 10 9 ). 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.

The DARPin Encyclopedia: from Basic Research towards Therapeutics

“…Notably, it has been demonstrated that recombinant VLR antibodies ('lampribodies') are amenable to selection by phage, yeast and human cell line display techniques, either from synthetic DNA libraries (designed VLRs) or from cloned VLR cDNAs isolated from lamprey larvae after immunization with an antigen. 30,31 Apart from their role in the adaptive immune system of jawless vertebrates, LRR motifs were found to play a vital role in the innate immune system of plants, invertebrates and vertebrates as part of pattern recognition receptors (e.g. Toll-like receptors) that are present at the cell surface or in endosomes detecting non-self structures from pathogens (e.g.…”

Biomolecules from Different Angles

“…Therefore, this Special Issue is a snapshot of a collective view of Biomolecules on biomolecules. The covered topics range from the description of secondary metabolites produced by fungi [ 1 ] to the overview of the phytochemical composition and pharmacological and food applications of Prosopis plants [ 2 ], to comparative evaluation of the antiproliferative activity of plant-derived silver nanoparticles synthesized using callus and anthocyanin extracts of purple basil (BC-AgNPs and AE-AgNPs, respectively) [ 3 ], to systematic description of the utilization of natural products as a source of antitumor drugs [ 4 ], to the analysis of the effects of caffeine and other methylxanthines on Aβ-homeostasis by shifting the α- and β-secretase-based processing of amyloid precursor protein (APP) from the Aβ-producing amyloidogenic to the non-amyloidogenic pathway [ 5 ], to the topoisomerase I and radical trapping antioxidant activities of a series of N-alkyl-acridones and N,N′-dialkyl-9,9′-biacridylidenes [ 6 ], to the analysis of the effects of phytoalexins in soybeans via synergistic inhibition on α-glucosidase [ 7 ], to functional and structural characterization of several important proteins (such as the analysis of the general and genomic DNA-binding specificity of a transcription factor TTHB023 from Thermus thermophilus HB8 [ 8 ], investigation of the voltage sensing in protein translocation via the bacterial channel SecYEG [ 9 ], and structural characterization of the terminase (packaging protein) pUL15, which is the most conserved among all the Herpes Simplex Virus 1 (HSV1) gene products [ 10 ]), to the description of utilization of kinetic transition in amyloid assembly for screening fluorophores for preferential responses to oligomer over fibril formation [ 11 ], to the description of the utilization of amyloid fibril biomaterials, designer amyloid cell-penetrating peptides comprised of β-sheet cores derived from naturally occurring protein sequences and designed positively charged and aromatic residues exposed at key residue positions as gene transfer vehicles capable of self-assembling and promoting the DNA condensation and cell internalization [ 12 ], to the introduction of the use of phage display system for generation of lamprey monoclonal antibodies, lampribodies, which are the variable lymphocyte receptors (VLRs) consisting of leucine rich repeats (LRRs), whose diversity is generated by stepwise genomic rearrangements of LRR cassettes dispersed throughout the VLRB locus [ 13 ], to the description of changes in the N-glycomic profile associated with post-traumatic stress disorder (PTSD) via comparative analysis of the N-glycomic profiles in 543 male Caucasian individuals (299 veterans with PTSD and 244 control subjects) [ 14 ], to the analysis of the effect of macromolecular crowding on the functionality of tumor suppressors ING containing a plant homeodomain (PHD) and their ability to interact with the histone H3 trimethylated at lysine 4 (H3K4me3) [ 15 ], and to the compariso...…”