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Proc Natl Acad Sci U S A. Jun 1984; 81(12): 3675–3679.
PMCID: PMC345281

Method to map antigenic determinants recognized by monoclonal antibodies: localization of a determinant of virus neutralization on the feline leukemia virus envelope protein gp70.


A method is presented whereby antigenic determinants recognized by specific monoclonal antibodies can be mapped to specific sites on a protein sequence with high resolution. Short DNase I-generated DNA fragments encoding portions of the protein of interest are molecularly cloned into the EcoRI site of the beta-galactosidase gene of phage lambda Charon 16 so as to obtain expression of random protein fragments as fusion proteins. The monoclonal antibody is used to screen the phage library to isolate phage expressing the specific antigenic determinant. DNA of immunoreactive phage can be analyzed rapidly and subcloned to allow DNA sequence determination. The method is generally applicable and permits antigenic determinants of functionally interesting monoclonal antibodies to be mapped and related to specific protein sequences. We have used this procedure to determine the region of the feline leukemia virus envelope protein gp70 recognized by a virus-neutralizing monoclonal antibody, cl.25. Antibody binding was mapped to a 14-amino acid region in the amino-terminal half of gp70. This region may be directly involved in an essential function of the gp70 protein, perhaps in gp70-mediated host recognition functions. Synthetic peptides derived from this region may provide useful vaccine antigens for the prevention of feline leukemia virus-associated disease in cats.

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  • Dietzschold B, Wiktor TJ, Macfarlan R, Varrichio A. Antigenic structure of rabies virus glycoprotein: ordering and immunological characterization of the large CNBr cleavage fragments. J Virol. 1982 Nov;44(2):595–602. [PMC free article] [PubMed]
  • Bachrach HL, Morgan DO, McKercher PD, Moore DM, Robertson BH. Foot-and-mouth disease virus: immunogenicity and structure of fragments derived from capsid protein VP and of virus containing cleaved VP. Vet Microbiol. 1982 May;7(2):85–96. [PubMed]
  • Jackson DC, Dopheide TA, Russell RJ, White DO, Ward CW. Antigenic determinants of influenza virus hemagglutinin. II. Antigenic reactivity of the isolated N-terminal cyanogen bromide peptide of A/Memphis/72 hemagglutinin heavy chain. Virology. 1979 Mar;93(2):458–465. [PubMed]
  • Cheung A, DeLamarter J, Weiss S, Küpper H. Comparison of the major antigenic determinants of different serotypes of foot-and-mouth disease virus. J Virol. 1983 Nov;48(2):451–459. [PMC free article] [PubMed]
  • Webster RG, Laver WG, Air GM, Schild GC. Molecular mechanisms of variation in influenza viruses. Nature. 1982 Mar 11;296(5853):115–121. [PubMed]
  • Caton AJ, Brownlee GG, Yewdell JW, Gerhard W. The antigenic structure of the influenza virus A/PR/8/34 hemagglutinin (H1 subtype). Cell. 1982 Dec;31(2 Pt 1):417–427. [PubMed]
  • Minor PD, Schild GC, Bootman J, Evans DM, Ferguson M, Reeve P, Spitz M, Stanway G, Cann AJ, Hauptmann R, et al. Location and primary structure of a major antigenic site for poliovirus neutralization. Nature. 1983 Feb 24;301(5902):674–679. [PubMed]
  • Evans DM, Minor PD, Schild GS, Almond JW. Critical role of an eight-amino acid sequence of VP1 in neutralization of poliovirus type 3. Nature. 1983 Aug 4;304(5925):459–462. [PubMed]
  • Bittle JL, Houghten RA, Alexander H, Shinnick TM, Sutcliffe JG, Lerner RA, Rowlands DJ, Brown F. Protection against foot-and-mouth disease by immunization with a chemically synthesized peptide predicted from the viral nucleotide sequence. Nature. 1982 Jul 1;298(5869):30–33. [PubMed]
  • Erlich HA, Cohen SN, McDevitt HO. A sensitive radioimmunoassay for detecting products translated from cloned DNA fragments. Cell. 1978 Apr;13(4):681–689. [PubMed]
  • Broome S, Gilbert W. Immunological screening method to detect specific translation products. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2746–2749. [PMC free article] [PubMed]
  • Hitzeman RA, Clarke L, Carbon J. Isolation and characterization of the yeast 3-phosphoglycerokinase gene (PGK) by an immunological screening technique. J Biol Chem. 1980 Dec 25;255(24):12073–12080. [PubMed]
  • Young RA, Davis RW. Yeast RNA polymerase II genes: isolation with antibody probes. Science. 1983 Nov 18;222(4625):778–782. [PubMed]
  • Rüther U, Koenen M, Sippel AE, Müller-Hill B. Exon cloning: immunoenzymatic identification of exons of the chicken lysozyme gene. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6852–6855. [PMC free article] [PubMed]
  • Hardy WD, Jr, Old LJ, Hess PW, Essex M, Cotter S. Horizontal transmission of feline leukaemia virus. Nature. 1973 Aug 3;244(5414):266–269. [PubMed]
  • Jarrett O, Hardy WD, Jr, Golder MC, Hay D. The frequency of occurrence of feline leukaemia virus subgroups in cats. Int J Cancer. 1978 Mar 15;21(3):334–337. [PubMed]
  • Pedersen NC, Theilen G, Keane MA, Fairbanks L, Mason T, Orser B, Che CH, Allison C. Studies of naturally transmitted feline leukemia virus infection. Am J Vet Res. 1977 Oct;38(10):1523–1531. [PubMed]
  • Sarma PS, Log T. Subgroup classification of feline leukemia and sarcoma viruses by viral interference and neutralization tests. Virology. 1973 Jul;54(1):160–169. [PubMed]
  • Sarma PS, Log T, Jain D, Hill PR, Huebner RJ. Differential host range of viruses of feline leukemia-sarcoma complex. Virology. 1975 Apr;64(2):438–446. [PubMed]
  • Russell PH, Jarrett O. The specificity of neutralizing antibodies to feline leukaemia viruses. Int J Cancer. 1978 Jun 15;21(6):768–778. [PubMed]
  • de Noronha F, Schäfer W, Essex M, Bolognesi DP. Influence of antisera to oncornavirus glycoprotein (gp71) on infections of cats with feline leukemia virus. Virology. 1978 Apr;85(2):617–621. [PubMed]
  • Schneider J, Falk H, Hunsmann G. Envelope polypeptides of Friend leukemia virus: purification and structural analysis. J Virol. 1980 Feb;33(2):597–605. [PMC free article] [PubMed]
  • Lutz H, Pedersen NC, Durbin R, Theilen GH. Monoclonal antibodies to three epitopic regions of feline leukemia virus p27 and their use in enzyme-linked immunosorbent assay of p27. J Immunol Methods. 1983 Jan 28;56(2):209–220. [PubMed]
  • Fischinger PJ, Blevins CS, Nomura S. Simple, quantitative assay for both xenotropic murine leukemia and ecotropic feline leukemia viruses. J Virol. 1974 Jul;14(1):177–179. [PMC free article] [PubMed]
  • Mullins JI, Casey JW, Nicolson MO, Burck KB, Davidson N. Sequence arrangement and biological activity of cloned feline leukemia virus proviruses from a virus-productive human cell line. J Virol. 1981 May;38(2):688–703. [PMC free article] [PubMed]
  • Blattner FR, Williams BG, Blechl AE, Denniston-Thompson K, Faber HE, Furlong L, Grunwald DJ, Kiefer DO, Moore DD, Schumm JW, et al. Charon phages: safer derivatives of bacteriophage lambda for DNA cloning. Science. 1977 Apr 8;196(4286):161–169. [PubMed]
  • Crouse GF, Simonsen CC, McEwan RN, Schimke RT. Structure of amplified normal and variant dihydrofolate reductase genes in mouse sarcoma S180 cells. J Biol Chem. 1982 Jul 10;257(13):7887–7897. [PubMed]
  • Franklin RM. Purification and properties of the replicative intermediate of the RNA bacteriophage R17. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1504–1511. [PMC free article] [PubMed]
  • Maniatis T, Hardison RC, Lacy E, Lauer J, O'Connell C, Quon D, Sim GK, Efstratiadis A. The isolation of structural genes from libraries of eucaryotic DNA. Cell. 1978 Oct;15(2):687–701. [PubMed]
  • Sternberg N, Tiemeier D, Enquist L. In vitro packaging of a lambda Dam vector containing EcoRI DNA fragments of Escherichia coli and phage P1. Gene. 1977 May;1(3-4):255–280. [PubMed]
  • Casadaban MJ, Cohen SN. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. [PubMed]
  • Young RA, Davis RW. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. [PMC free article] [PubMed]
  • Vieira J, Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. [PubMed]
  • Maxam AM, Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. [PubMed]
  • Kalnins A, Otto K, Rüther U, Müller-Hill B. Sequence of the lacZ gene of Escherichia coli. EMBO J. 1983;2(4):593–597. [PMC free article] [PubMed]
  • Büchel DE, Gronenborn B, Müller-Hill B. Sequence of the lactose permease gene. Nature. 1980 Feb 7;283(5747):541–545. [PubMed]
  • Erlich HA, Rodgers G, Vaillancourt P, Araujo FG, Remington JS. Identification of an antigen-specific immunoglobulin M antibody associated with acute Toxoplasma infection. Infect Immun. 1983 Aug;41(2):683–690. [PMC free article] [PubMed]
  • Rasheed S, Gardner MB. Isolation of feline leukemia virus from a leopard cat cell line and search for retrovirus in wild felidae. J Natl Cancer Inst. 1981 Oct;67(4):929–933. [PubMed]
  • Benton WD, Davis RW. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. [PubMed]
  • Elder JH, Mullins JI. Nucleotide sequence of the envelope gene of Gardner-Arnstein feline leukemia virus B reveals unique sequence homologies with a murine mink cell focus-forming virus. J Virol. 1983 Jun;46(3):871–880. [PMC free article] [PubMed]
  • Nunberg JH, Williams ME, Innis MA. Nucleotide sequences of the envelope genes of two isolates of feline leukemia virus subgroup B. J Virol. 1984 Feb;49(2):629–632. [PMC free article] [PubMed]
  • Messing J, Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. [PubMed]
  • Sanger F, Coulson AR, Barrell BG, Smith AJ, Roe BA. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. [PubMed]
  • Heidecker G, Messing J, Gronenborn B. A versatile primer for DNA sequencing in the M13mp2 cloning system. Gene. 1980 Jun;10(1):69–73. [PubMed]
  • Kleid DG, Yansura D, Small B, Dowbenko D, Moore DM, Grubman MJ, McKercher PD, Morgan DO, Robertson BH, Bachrach HL. Cloned viral protein vaccine for foot-and-mouth disease: responses in cattle and swine. Science. 1981 Dec 4;214(4525):1125–1129. [PubMed]
  • van der Werf S, Wychowski C, Bruneau P, Blondel B, Crainic R, Horodniceanu F, Girard M. Localization of a poliovirus type 1 neutralization epitope in viral capsid polypeptide VP1. Proc Natl Acad Sci U S A. 1983 Aug;80(16):5080–5084. [PMC free article] [PubMed]
  • Wychowski C, van der Werf S, Siffert O, Crainic R, Bruneau P, Girard M. A poliovirus type 1 neutralization epitope is located within amino acid residues 93 to 104 of viral capsid polypeptide VP1. EMBO J. 1983;2(11):2019–2024. [PMC free article] [PubMed]
  • Bosselman RA, van Straaten F, Van Beveren C, Verma IM, Vogt M. Analysis of the env gene of a molecularly cloned and biologically active Moloney mink cell focus-forming proviral DNA. J Virol. 1982 Oct;44(1):19–31. [PMC free article] [PubMed]
  • Shinnick TM, Lerner RA, Sutcliffe JG. Nucleotide sequence of Moloney murine leukaemia virus. Nature. 1981 Oct 15;293(5833):543–548. [PubMed]
  • Koch W, Hunsmann G, Friedrich R. Nucleotide sequence of the envelope gene of Friend murine leukemia virus. J Virol. 1983 Jan;45(1):1–9. [PMC free article] [PubMed]
  • Lenz J, Crowther R, Straceski A, Haseltine W. Nucleotide sequence of the Akv env gene. J Virol. 1982 May;42(2):519–529. [PMC free article] [PubMed]
  • Chen R. Complete amino acid sequence and glycosylation sites of glycoprotein gp71A of Friend murine leukemia virus. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5788–5792. [PMC free article] [PubMed]
  • Hopp TP, Woods KR. Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824–3828. [PMC free article] [PubMed]

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