+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Two antipeptide monoclonal antibodies that recognize adhesive sequences in fibrinogen: identification of antigenic determinants and unrelated sequences using synthetic combinatorial libraries

Two antipeptide monoclonal antibodies that recognize adhesive sequences in fibrinogen: identification of antigenic determinants and unrelated sequences using synthetic combinatorial libraries

Biomedical Peptides Proteins and Nucleic Acids 1(3): 199-204

The fine specificity of two different monoclonal antibodies raised against synthetic peptides, each representing one of the two Arg-Gly-Asp (RGD) sequences in fibrinogen, was examined using synthetic combinatorial libraries (SCLs). The monoclonal antibodies (mAb), mAb LJ-134B/29 and mAb LJ-155B/16, recognize both the immunogenic peptide and native fibrinogen. The specificity of mAb LJ-134B29 was mapped using hexa- and decapeptide positional scanning SCLs (PS-SCLs) and competitive ELISA. The most active amino acids at each position of the two libraries were identified from a single screening. Individual hexa- and decapeptides were synthesized and assayed to determine their binding affinities. The 16 individual hexapeptides represented single and multiple substitutions of the antigenic determinant sequence, -GDSTFE-, eight of which had affinities less than 10nM. Four of the twelve individual decapeptides were found to have binding affinities of approximately 300nM, or nearly three-fold less than the peptide immunogen. A dual-defined hexapeptide library was screened against mAb LJ-155B/16, and individual peptides were obtained through an iterative selection and synthesis process. Surprisingly, one of the most active sequences was Ac-WWYESW-NH2 (IC50 = 40nM), which showed no similarity to the sequence of the immunizing peptide. Further mapping of the specificity of this antibody revealed that the antigenic determinant within the peptide immunogen was not completely linear. Recognition of this unrelated sequence by mAb LJ-155B/16 was confirmed in a direct binding assay using biotinylated peptide. The use of SCLs for the elucidation of high affinity peptides recognized by these two antibodies may provide additional information on the molecular mechanisms of fibrinogen binding to different integrin receptors.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 047853753

Download citation: RISBibTeXText

PMID: 9346853

Related references

Synthetic peptide combinatorial libraries. Bio/Technology 12(7): 709-710, 1994

Short exposure to Notch ligand Delta-4 is sufficient to induce T-cell differentiation program and to increase the T cell potential of primary human CD34+ cells. Experimental Hematology 34(12): 1720-1729, 2006

Antipeptide antibodies differentiate between long and short isoforms of the D2 dopamine receptor. Biochemical and Biophysical Research Communications 179(2): 824-829, 1991

Probing the sensitivity of phosphorus 31 nmr chemical shifts to hydrogen bonding and to stereochemistry. Febs Letters 105(1): 11-14, 1979

Tea asthma response to specific and non specific challenges. British Journal Of Industrial Medicine. 46(5): 350-351, 1989

HLA-E molecule induction on the surface of tumor cells protects them from cytotoxic lymphocytes. Voprosy Onkologii 55(2): 224-229, 2009

Monoclonal antibodies to thyroxine t4 production and characterization. Journal of Endocrinology 112(Suppl.), 1987

Monoclonal antipeptide antibodies against amino acid residues 1101-1106 of human C4 distinguish C4A from C4B. Complement and Inflammation 8(1): 33-42, 1991

Amino acid sequences of a heterodimeric neurotoxin from the venom of the false horned viper. Toxicon 33(7): 863-874, 1995

Identification of antigenic escape variants in an immunodominant epitope of hepatitis C virus. International Immunology 11(4): 577-583, 1999

Antigenic determinants of influenza virus hemagglutinin. XII. the epitopes of a synthetic peptide representing the C-terminus of HA1. Virology 155(2): 625-632, 1986

A species-specific amino acid difference in the macaque CD4 receptor restricts replication by global circulating HIV-1 variants representing viruses from recent infection. Journal of Virology 86(23): 12472-12483, 2012

Monoclonal antibodies to Chlamydia trachomatis: antibody specificities and antigen characterization. Journal of Immunology 128(3): 1083-1089, 1982

Two DNA restriction fragment length polymorphisms associated with Ag(t/z) and Ag(g/c) antigenic sites of human apolipoprotein B. Arteriosclerosis 7(3): 301-305, 1987

Monoclonal antipeptide antibodies recognize IL-3 and neutralize its bioactivity in vivo. Journal of Immunology 140(4): 1182-1187, 1988