Specificities of broadly neutralizing anti-HIV-1 sera

Current Opinion in HIV and AIDS

Volumn 4, Issue 5, 2009, Pages 364-372

  Binley, James ^a  

^a TORREY PINES INSTITUTE FOR MOLECULAR STUDIES   (United States)

Author keywords

gp120; gp41; HIV 1; Neutralization

Indexed keywords

EPITOPE; GLYCOPROTEIN GP 120; HUMAN IMMUNODEFICIENCY VIRUS ANTIBODY; HUMAN IMMUNODEFICIENCY VIRUS VACCINE; MONOCLONAL ANTIBODY; NEUTRALIZING ANTIBODY; PEPTIDE; VIRUS PROTEIN;

ANTIBODY SPECIFICITY; DISEASE CLASSIFICATION; GENE MAPPING; HUMAN; HUMAN IMMUNODEFICIENCY VIRUS 1 INFECTION; HUMAN IMMUNODEFICIENCY VIRUS 2; IMMUNOGENICITY; NONHUMAN; POLYACRYLAMIDE GEL ELECTROPHORESIS; PREVALENCE; PRIORITY JOURNAL; PROTEIN ANALYSIS; REVIEW; SERUM; SIMIAN IMMUNODEFICIENCY VIRUS; SPIKE; VIRUS ENVELOPE; VIRUS NEUTRALIZATION;

ANTIBODIES, NEUTRALIZING; CROSS REACTIONS; EPITOPE MAPPING; HIV ANTIBODIES; HIV INFECTIONS; HIV-1; HUMANS; SERUM;

EID: 69549086342     PISSN: 1746630X     EISSN: 17466318     Source Type: Journal    
DOI: 10.1097/COH.0b013e32832e06fe     Document Type: Review

References (91)

1. Kraft Z, Derby NR, McCaffrey RA, et al. Macaques infected with a CCR5-tropic simian/human immunodeficiency virus (SHIV) develop broadly reactive anti-HIV neutralizing antibodies. J Virol 2007; 81:6402-6411. (Pubitemid 46878046)
2. Li Y, Migueles S, Welcher B, et al. Broad HIV-1 neutralization mediated by CD4 binding site antibodies. Nat Med 2007; 13:1032-1034. This paper describes the first detailed attempt to map the specificities responsible for the broad neutralizing activities of two chronically infected slow progressing patients. Importantly, both these initial patients appeared to make b12-like CD4bs overlapping nAbs. (Pubitemid 47517505)
3. Santiago ML, Montano M, Benitez R, et al. Apobec3 encodes Rfv3, a gene influencing neutralizing antibody control of retrovirus infection. Science 2008; 321:1343-1346.
4. Kostrikis LG, Cao Y, Ngai H, et al. Quantitative analysis of serum neutralization of human immunodeficiency virus type 1 from subtypes A, B, C, D, E, F, and I: lack of direct correlation between neutralization serotypes and genetic subtypes and evidence for prevalent serum-dependent infectivity enhancement. J Virol 1996; 70:445-458.
5. Brown BK, Wieczorek L, Sanders-Buell E, et al. Cross-clade neutralization patterns among HIV-1 strains from the six major clades of the pandemic evaluated and compared in two different models. Virology 2008; 375:529-538. The intra- and interclade neutralizing potency of plasma pools from various clades was evaluated in pseudovirus and peripheral blood mononuclear blood cell (PBMC) blast neutralization assays.
6. Rademeyer C, Moore PL, Taylor N, et al. Genetic characteristics of HIV-1 subtype C envelopes inducing cross-neutralizing antibodies. Virology 2007; 368:172-181.
7. Moore JP, Cao Y, Leu J, et al. Inter- and intraclade neutralization of human immunodeficiency virus type 1: genetic clades do not correspond to neutralization serotypes but partially correspond to gp120 antigenic serotypes. J Virol 1996; 70:427-444.
8. Pantophlet R, Burton DR. GP120: target for neutralizing HIV-1 antibodies. Annu Rev Immunol 2006; 24:739-769.
9. Sather DN, Armann J, Ching LK, et al. Factors associated with the development of cross-reactive neutralizing antibodies during HIV-1 infection. J Virol 2008; 83:757-769. Time since infection and ongoing viral replication were found to correlate with the development of broad neutralizing responses. The specificities of the broad sera were variably adsorbed by gp120 and a fraction was specific for the CD4bs epitope. Significant MPER activity was noted in a plasma with moderate breadth.
10. Binley JM, Wrin T, Korber B, et al. Comprehensive cross-clade neutralization analysis of a panel of antihuman immunodeficiency virus type 1 monoclonal antibodies. J Virol 2004; 78:13232-13252.
11. Montefiori D, Sattentau Q, Flores J, et al. Antibody-based HIV-1 vaccines: recent developments and future directions. PLoS Med 2007; 4:e348.
12. Simek MD, Rida W, Priddy F, et al. HIV-1 elite neutralizers: individuals with broad and potent neutralizing activity identified using high throughput neutralization assay together with an analytical selection algorithm. J Virol 2009; 83:7337-7348. A survey of large cohorts of serumsamples in neutralization assays provided information on the frequency of broad neutralization and identified elite neutralizers of particular interest for attempts to map broad neutralization and to isolate new bnmAbs.
13. Stamatatos L, Morris L, Burton DR, Mascola J. Neutralizing antibodies generated during natural hiv-1 infection: good news for an HIV-1 vaccine? Nat Med 2009 Jun 14. [Epub ahead of print]. The wide gap in efficacy and breadth of HIV-1 neutralizing responses generated in natural infection and by candidate vaccines is discussed, and suggestions as to how this gap may be bridged are put forward.
14. Haynes BF, Montefiori DC. Aiming to induce broadly reactive neutralizing antibody responses with HIV-1 vaccine candidates. Expert Rev Vaccin 2006; 5:347-363.
15. Phogat S, Wyatt RT, Karlsson Hedestam GB. Inhibition of HIV-1 entry by antibodies: potential viral and cellular targets. J Intern Med 2007; 262:26-43.
16. Fouts TR, Binley JM, Trkola A, et al. Neutralization of the human immunodeficiency virus type 1 primary isolate JR-FL by human monoclonal antibodies correlates with antibody binding to the oligomeric form of the envelope glycoprotein complex. J Virol 1997; 71:2779-2785.
17. Moore PL, Crooks ET, Porter L, et al. Nature of nonfunctional envelope proteins on the surface of human immunodeficiency virus type 1. J Virol 2006; 80:2515-2528.
18. Sattentau QJ, Moore JP. Human immunodeficiency virus type 1 neutralization is determined by epitope exposure on the gp120 oligomer. J Exp Med 1995; 182:185-196.
19. Burton DR, Pyati J, Koduri R, et al. Efficient neutralization of primary isolates of HIV-1 by a recombinant human monoclonal antibody. Science 1994; 266:1024-1027.
20. Calarese DA, Scanlan CN, Zwick MB, et al. Antibody domain exchange is an immunological solution to carbohydrate cluster recognition. Science 2003; 300:2065-2071.
21. Trkola A, Purtscher M, Muster T, et al. Human monoclonal antibody 2G12 defines a distinctive neutralization epitope on the gp120 glycoprotein of human immunodeficiency virus type 1. J Virol 1996; 70:1100-1108.
22. Scanlan CN, Pantophlet R, Wormald MR, et al. The carbohydrate epitope of the neutralizing anti-HIV-1 antibody 2G12. Adv Exp Med Biol 2003; 535:205-218.
23. Scanlan CN, Ritchie GE, Baruah K, et al. Inhibition of mammalian glycan biosynthesis produces nonself antigens for a broadly neutralizing, HIV-1 specific antibody. J Mol Biol 2007; 372:16-22.
24. Sanders RW, Venturi M, Schiffner L, et al. The mannose-dependent epitope for neutralizing antibody 2G12 on human immunodeficiency virus type 1 glycoprotein gp120. J Virol 2002; 76:7293-7305.
25. Nelson JD, Brunel FM, Jensen R, et al. An affinity-enhanced neutralizing antibody against the membrane-proximal external region of human immunodeficiency virus type 1 gp41 recognizes an epitope between those of 2F5 and 4E10. J Virol 2007; 81:4033-4043.
26. Mascola JR, Lewis MG, Stiegler G, et al. Protection of macaques against pathogenic simian/human immunodeficiency virus 89.6PD by passive transfer of neutralizing antibodies. J Virol 1999; 73:4009-4018.
27. Mascola JR, Stiegler G, VanCott TC, et al. Protection of macaques against vaginal transmission of a pathogenic HIV-1/SIV chimeric virus by passive infusion of neutralizing antibodies. Nat Med 2000; 6:207-210.
28. Mascola JR. Passive transfer studies to elucidate the role of antibody-mediated protection against HIV-1. Vaccine 2002; 20:1922-1925.
29. Baba TW, Liska V, Hofmann-Lehmann R, et al. Human neutralizing monoclonal antibodies of the IgG1 subtype protect against mucosal simian-human immunodeficiency virus infection. Nat Med 2000; 6:200-206.
30. Parren PW, Marx PA, Hessell AJ, et al. Antibody protects macaques against vaginal challenge with a pathogenic R5 simian/human immunodeficiency virus at serum levels giving complete neutralization in vitro. J Virol 2001; 75:8340-8347.
31. Hessell AJ, Hangartner L, Hunter M, et al. Effective, low titer, antibody protection against low dose repeated mucosal challenge in macaques. Nat Med 2009 Jun 7. [Epub ahead of print].
32. Nishimura Y, Igarashi T, Haigwood NL, et al. Transfer of neutralizing IgG to macaques 6 h but not 24 h after SHIV infection confers sterilizing protection: implications for HIV-1 vaccine development. Proc Natl Acad Sci U S A 2003; 100:15131-15136.
33. Binley JM, Lybarger EA, Crooks ET, et al. Profiling the specificity of neutralizing antibodies in a large panel of HIV-1 plasmas from subtype B and C chronic infections. J Virol 2008; 82:11651-11668. In this collaborative study, a panel 24 clade B and C broadly neutralizing plasmas were mapped using multiple contemporary methods. Some plasmas recognized the CD4bs and one recognized the MPER region, but 2/3 of the activity could not be mapped to any known epitope.
34. Dhillon AK, Donners H, Pantophlet R, et al. Dissecting the neutralizing antibody specificities of broadly neutralizing sera from human immunodeficiency virus type 1-infected donors. J Virol 2007; 81:6548-6562. Using gp120 fractionation to analyze three broadly neutralizing sera, it was found that the efficiency of fractionation depended on the isolate against which neutralization was measured and the nature of the gp120 protein. A core gp120 was often inefficient at adsorbing neutralization, suggesting somerole of variable loop domains.
35. Li Y, Svehla K, Louder MK, et al. Analysis of neutralization specificities in polyclonal sera derived from human immunodeficiency virus type 1-infected individuals. J Virol 2009; 83:1045-1059. In a systematic follow-up to their initial gp120 fractionation study, further evidence for CD4bs nAbs was presented, as was evidence of a contribution of CD4i nAbs. 4E10-like MPER nAbs constitute a significant fraction of neutralizing activity against a primary isolate in one serum.
36. Gray ES, Taylor N, Wycuff D, et al. Antibody specificities associated with neutralization breadth in plasma from HIV-1 subtype C infected blood donors. J Virol 2009 Jun 24. [Epub ahead of print]. The neutralizing activity of clade C plasmas was investigated using clade C reagents. Some gp120-specific activity was found, but this mostly did not target the D368R D368R-sensitive CD4bs epitope. A correlation between the detection of CD4i, MPER and autoimmune responses and neutralization breadth was noted, although it is unclear whether these factors play a direct role or are merely surrogate markers.
37. Binley JM, Cayanan CS, Wiley C, et al. Redox-triggered infection by disulfide-shackled human immunodeficiency virus type 1 pseudovirions. J Virol 2003; 77:5678-5684.
38. Abrahamyan LG, Markosyan RM, Moore JP, et al. Human immunodeficiency virus type 1 Env with an intersubunit disulfide bond engages coreceptors but requires bond reduction after engagement to induce fusion. J Virol 2003; 77:5829-5836.
39. Crooks ET, Moore PL, Richman D, et al. Characterizing anti-HIV monoclonal antibodies and immune sera by defining the mechanism of neutralization. Hum Antibodies 2005; 14 (3-4):101-113.
40. Decker JM, Bibollet-Ruche F, Wei X, et al. Antigenic conservation and immunogenicity of the HIV coreceptor binding site. J Exp Med 2005; 201:1407-1419.
41. Davis KL, Bibollet-Ruche F, Li H, et al. HIV-2/HIV-1 envelope chimeras detect high titers of HIV-1 V3-specific antibodies in human plasma. J Virol 2008; 83:1240-1259. An HIV-2 Env backbone was used as a host for HIV-1 V3 sequences in the construction of novel chimeric viruses for detecting V3 Abs.
42. Gray ES, Moore PL, Bibollet-Ruche F, et al. 4E10-resistant variants in a human immunodeficiency virus type 1 subtype C-infected individual with an anti-membrane-proximal external region-neutralizing antibody response. J Virol 2008; 82:2367-2375. This study describes a 4E10-resistant virus observed in a perinatally infected child that was apparently generated as a result of immune escape from nAb responses.
43. Gray ES, Moore PL, Choge IA, et al. Neutralizing antibody responses in acute human immunodeficiency virus type 1 subtype C infection. J Virol 2007; 81:6187-6196.
44. Yuste E, Sanford HB, Carmody J, et al. Simian immunodeficiency virus engrafted with human immunodeficiency virus type 1 (HIV-1)-specific epitopes: replication, neutralization, and survey of HIV-1-positive plasma. J Virol 2006; 80:3030-3041.
45. Li Y, Svehla K, Mathy NL, et al. Characterization of antibody responses elicited by human immunodeficiency virus type 1 primary isolate trimeric and monomeric envelope glycoproteins in selected adjuvants. J Virol 2006; 80:1414-1426.
46. SF162 and heterologous HIV-1 infection. J Virol 2006; 80:8745-8762.
47. Bibollet-Ruche F, Hui L, Decker JM, et al. Detection of novel neutralizing antibody responses to the membrane proximal external region (MPER) or gp41 following infection by HIV-1 subtypes A, B, C, D, F, G, H, CRF01, CRF02, or CRF11 [abstract #110]. Keystone Symposium X6, HIV vaccines . Keystone, CO; 27 March to 2 April 2006.
48. Ching LK, Vlachogiannis G, Bosch KA, Stamatatos L. The first hypervariable region of the gp120 Env glycoprotein defines the neutralizing susceptibility of heterologous human immunodeficiency virus type 1 isolates to neutralizing antibodies elicited by the SF162 gp140 immunogen. J Virol 2008; 82:949-956.
49. Pinter A, Honnen WJ, He Y, et al. The V1/V2 domain of gp120 is a global regulator of the sensitivity of primary human immunodeficiency virus type 1 isolates to neutralization by antibodies commonly induced upon infection. J Virol 2004; 78:5205-5215.
50. Davis KL, Bibollet-Ruche F, Li H, et al. Human immunodeficiency virus type 2 (HIV-2)/HIV-1 envelope chimeras detect high titers of broadly reactive HIV-1 V3-specific antibodies in human plasma. J Virol 2009; 83:1240-1259.
51. Pantophlet R, Ollmann Saphire E, Poignard P, et al. Fine mapping of the interaction of neutralizing and nonneutralizing monoclonal antibodies with the CD4 binding site of human immunodeficiency virus type 1 gp120. J Virol 2003; 77:642-658.
52. Scanlan CN, Pantophlet R, Wormald MR, et al. The broadly neutralizing antihuman immunodeficiency virus type 1 antibody 2G12 recognizes a cluster of a1->2 mannose residues on the outer face of gp120. J Virol 2002; 76:7306-7321.
53. Berman P, Schweighardt B, O'Rourke S, et al. Use of intra-patient sequence variation in viruses from recent infections to characterize epitopes recognized by broadly neutralizing antibodies [abstract P04-07 2008]. AIDS Vaccine 08 Conference. Cape Town, South Africa; 13-16 October. A mapping approach wherein the Env sequences of neutralization-susceptible versus resistant viruses were identified, followed by mutagenesis to identify the specific residue(s) that correlate with resistance.
54. Poignard P, Sabbe R, Picchio GR, et al. Neutralizing antibodies have limited effects on the control of established HIV-1 infection in vivo. Immunity 1999; 10:431-438.
55. Trkola A, Kuster H, Rusert P, et al. Delay of HIV-1 rebound after cessation of antiretroviral therapy through passive transfer of human neutralizing antibodies. Nat Med 2005; 11:615-622.
56. Crooks ET, Jiang P, Franti M, et al. Relationship of HIV-1 and SIV envelope glycoprotein trimer occupation and neutralization. Virology 2008; 377:364-378. The quantitative relationship between Env trimer binding and virus neutralization was explored.
57. Crooks ET, Moore PL, Franti M, et al. A comparative immunogenicity study of HIV-1 virus-like particles bearing various forms of envelope proteins, particles bearing no envelope and soluble monomeric gp120. Virology 2007; 366:245-262.
58. D'Amelio R, Biselli R, Nisini R, et al. Spectrotype of antigp120 antibodies remains stable during the course of HIV disease. J Acquir Immune Defic Syndr 1992; 5:930-935.
59. Barbas CF 3rd, Collet TA, Amberg W, et al. Molecular profile of an antibody response to HIV-1 as probed by combinatorial libraries. J Mol Biol 1993; 230:812-823.
60. Bowley DR, Labrijn AF, Zwick MB, Burton DR. Antigen selection from an HIV-1 immune antibody library displayed on yeast yields many novel antibodies compared to selection from the same library displayed on phage. Protein Eng Des Sel 2007; 20:81-90.
61. Moore PL, Ranchobe N, Lambson BE, et al. Analysis of neutralization escape suggests limited neutralizing antibody specificities in early HIV-1 subtype C infection [abstract OA03-04 2008]. AIDS Vaccine 2008. Cape Town, South Africa; 13-16 October 2008.
62. Scheid JF, Mouquet H, Feldhahn N, et al. Broad diversity of neutralizing antibodies isolated from memory B cells in HIV-infected individuals. Nature 2009; 458:636-640. Using single-cell PCR complementary DNA libraries cloned from memory B cells of infected donors, 134 mAb clones were isolated from six HIV-1-infected persons. None of these new mAbs were, however, broadly neutralizing, but in some cases exhibited strain-restricted activity against certain tier 2 viruses. It was suggested the broad activity of the parent sera may arise from the combined effects of multiple nAb specificities.
63. •]).
64. Shen X, Parks RJ, Montefiori DC, et al. In vivo gp41 antibodies targeting the 2F5 mAb epitope mediate HIV-1 neutralization breadth. J Virol 2009; 83:3617-3625. This is the first description of a patient who developed 2F5-like nAb responses. Interestingly, these responses developed in parallel with autoantibodies, consistent with the idea that MPER-specific nAbs may be regulated by tolerance.
65. Yang X, Tomov V, Kurteva S, et al. Characterization of the outer domain of the gp120 glycoprotein from human immunodeficiency virus type 1. J Virol 2004; 78:12975-12986.
66. Braibant M, Brunet S, Costagliola D, et al. Antibodies to conserved epitopes of the HIV-1 envelope in sera from long-term nonprogressors: prevalence and association with neutralizing activity. AIDS 2006; 20:1923-1930.
67. Tomaras GD, Yates NL, Liu P, et al. Initial B-cell responses to transmitted human immunodeficiency virus type 1: virion-binding immunoglobulin M (IgM) and IgG antibodies followed by plasma antigp41 antibodies with ineffective control of initial viremia. J Virol 2008; 82:12449-12463. The development of Ab responses was tracked before, during and after the initial ramp-up of viral load during acute infection, revealing an insight into the earliest immunodominant specificities generated, none of which are neutralizing.
68. Haynes BF, Fleming J, St Clair EW, et al. Cardiolipin polyspecific autoreactivity in two broadly neutralizing HIV-1 antibodies. Science 2005; 308:1906-1908.
69. Martinez V, Diemert MC, Braibant M, et al. Anticardiolipin antibodies in HIV infection are independently associated with antibodies to the membrane proximal external region of gp41 and with cell-associated HIV DNA and immune activation. Clin Infect Dis 2009; 48:123-132.
70. Bou-Habib DC, Roderiquez G, Oravecz T, et al. Cryptic nature of envelope V3 region epitopes protects primary monocytotropic human immunodeficiency virus type 1 from antibody neutralization. J Virol 1994; 68:6006-6013.
71. Krachmarov C, Pinter A, Honnen WJ, et al. Antibodies that are cross-reactive for human immunodeficiency virus type 1 clade A and clade B V3 domains are common in patient sera from Cameroon, but their neutralization activity is usually restricted by epitope masking. J Virol 2005; 79:780-790.
72. Krachmarov CP, Honnen WJ, Kayman SC, et al. Factors determining the breadth and potency of neutralization by V3-specific human monoclonal antibodies derived from subjects infected with clade A or clade B strains of human immunodeficiency virus type 1. J Virol 2006; 80:7127-7135.
73. Pantophlet R, Wrin T, Cavacini LA, et al. Neutralizing activity of antibodies to the V3 loop region of HIV-1 gp120 relative to their epitope fine specificity. Virology 2008; 381:251-260.
74. Labrijn AF, Poignard P, Raja A, et al. Access of antibody molecules to the conserved coreceptor binding site on glycoprotein gp120 Is sterically restricted on primary human immunodeficiency virus type 1. J Virol 2003; 77:10557-10565.
75. He Y, Honnen WJ, Krachmarov CP, et al. Efficient isolation of novel human monoclonal antibodies with neutralizing activity against HIV-1 from transgenic mice expressing human Ig loci. J Immunol 2002; 169:595-605.
76. Moore PL, Gray ES, Choge IA, et al. The C3-V4 region is a major target of autologous neutralizing antibodies in HIV-1 subtype C infection. J Virol 2007; 82:1860-1869.
77. Rong R, Bibollet-Ruche F, Mulenga J, et al. Role of V1V2 and other human immunodeficiency virus type 1 envelope domains in resistance to autologous neutralization during clade C infection. J Virol 2007; 81:1350-1359.
78. Nolan KM, Jordan AP, Hoxie JA. Effects of partial deletions within the human immunodeficiency virus type 1 V3 loop on coreceptor tropism and sensitivity to entry inhibitors. J Virol 2008; 82:664-673.
79. Cormier EG, Dragic T. The crown and stem of the V3 loop play distinct roles in human immunodeficiency virus type 1 envelope glycoprotein interactions with the CCR5 coreceptor. J Virol 2002; 76:8953-8957.
80. Suphaphiphat P, Essex M, Lee TH. Mutations in the V3 stem versus the V3 crown and C4 region have different effects on the binding and fusion steps of human immunodeficiency virus type 1 gp120 interaction with the CCR5 coreceptor. Virology 2007; 360:182-190.
81. Arthos J, Cicala C, Martinelli E, et al. HIV-1 envelope protein binds to and signals through integrin alpha4beta7, the gut mucosal homing receptor for peripheral T cells. Nat Immunol 2008; 9:301-309.
82. Gorny MK, Stamatatos L, Volsky B, et al. Identification of a new quaternary neutralizing epitope on human immunodeficiency virus type 1 virus particles. J Virol 2005; 79:5232-5237.
83. Robinson JE, Franco K, Elliott D, et al. Human and rhesus mAbs recognizing distinct groups of epitopes that elicit neutralizing antibodies against autologous HIV-1 strains [abstract PO4-18 2008]. AIDS Vaccine 08 Conference. Cape Town, South Africa; 13-16 October 2008. Neutralizing mAbs that target quaternary trimer-specific epitopes of the SF162 isolate were described.
84. Honnen WJ, Krachmarov C, Kayman SC, et al. Type-specific epitopes targeted by monoclonal antibodies with exceptionally potent neutralizing activities for selected strains of human immunodeficiency virus type 1 map to a common region of the V2 domain of gp120 and differ only at single positions from the clade B consensus sequence. J Virol 2007; 81:1424-1432.
85. Rong R, Gnanakaran S, Decker JM, et al. Unique mutational patterns in the envelope alpha 2 amphipathic helix and acquisition of length in gp120 hypervariable domains are associated with resistance to autologous neutralization of subtype C human immunodeficiency virus type 1. J Virol 2007; 81:5658-5668.
86. Wu L, Yang ZY, Xu L, et al. Cross-clade recognition and neutralization by the V3 region from clade C human immunodeficiency virus-1 envelope. Vaccine 2006; 24:4995-5002.
87. Vaine M, Wang S, Crooks ET, et al. Improved induction of antibodies against key neutralizing epitopes by HIV-1 gp120 DNA prime-protein boost vaccination compared to gp120 protein only vaccination. J Virol 2008; 82:7369-7378.
88. Tiller T, Meffre E, Yurasov S, et al. Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning. J Immunol Methods 2008; 329 (1-2):112-124.
89. Wrammert J, Smith K, Miller J, et al. Rapid cloning of high-affinity human monoclonal antibodies against influenza virus. Nature 2008; 453:667-671. A rapid method is described for cloning mAbs, sourcing from the high counts of circulating antibody-secreting cells generated by vaccination.
90. Traggiai E, Becker S, Subbarao K, et al. An efficient method to make human monoclonal antibodies from memory B cells: potent neutralization of SARS coronavirus. Nat Med 2004; 10:871-875.
91. Moody MA, Haynes BF. Antigen-specific B cell detection reagents: use and quality control. Cytometry A 2008; 73:1086-1092.