1887

Chapter 8 : Antigen-Antibody Reactions

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Ebook: Choose a downloadable PDF or ePub file. Chapter is a downloadable PDF file. File must be downloaded within 48 hours of purchase

Buy this Chapter
Digital (?) $30.00

Preview this chapter:
Zoom in
Zoomout

Antigen-Antibody Reactions, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555817497/9781555812232_Chap08-1.gif /docserver/preview/fulltext/10.1128/9781555817497/9781555812232_Chap08-2.gif

Abstract:

Reactions between antigens (Ags) and antibodies (Abs) are usefully exploited in many areas of life science research. The monoclonal Ab (MAb) technology developed by Köhler and Milstein allows for the production of unlimited quantities of Abs against virtually any molecule. As there is an immense volume of information concerning all aspects of Ag-Ab reactions in the literature, the objective of this chapter is to provide simple and useful protocols and an introduction to some of the more novel techniques. The routes of injection commonly used on rabbits, mice and rats include intradermal (i.d.), subcutaneous (s.c.), intramuscular (i.m.), intraperitoneal (i.p.), and intravenous (i.v.). Nucleic acid (DNA or RNA) immunization and genetic vaccines introduced in the early 1990s are some of the most important discoveries and novel strategies in vaccine development. The essential features of a DNA vaccine are a bacterial plasmid vector engineered to carry a DNA insert encoding the protein immunogen(s) of interest, a eukaryote gene promoter, and a poly(A) site to enable expression of the protein in mammalian cells. The vectors are usually maintained in and purified from . The polyclonal serum is expressed from the blood clot by centrifugation, and approximately 52% of its volume can be collected as serum. The serum can be used directly in Ag-Ab reactions, absorbed to deplete it of nonspecific or cross-reactive Abs, or fractionated to purify Abs free of other serum proteins.

Citation: Mutharia L, Lam J. 2007. Antigen-Antibody Reactions, p 138-167. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch08

Key Concept Ranking

Outer Membrane Proteins
0.44863775
Agarose Gel Electrophoresis
0.42233074
Chemicals
0.41793635
Enzyme-Linked Immunosorbent Assay
0.411093
0.44863775
Highlighted Text: Show | Hide
Loading full text...

Full text loading...

Figures

Image of FIGURE 1
FIGURE 1

HPLC separation of IgM MAb from ascites fluid. The first peak represents unbound material, and the second peak represents pure IgM Abs eluted after the addition of elution buffer 1. The arrow indicates the time at which the elution buffer was added.

Citation: Mutharia L, Lam J. 2007. Antigen-Antibody Reactions, p 138-167. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch08
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of FIGURE 2
FIGURE 2

SDS-PAGE profiles of IgM MAb purification by HPLC. Lanes: 1, molecular mass standards; 2, ascites fluid before purification (note the large number of bands that are normally found in animal serum proteins or body fluids); and 3, purified IgM MAb eluted from the column (the second major peak in Fig. 1 ). Note the presence of only two major bands representing the heavy and light chains at apparent molecular masses of 60 and 28 kDa, respectively.

Citation: Mutharia L, Lam J. 2007. Antigen-Antibody Reactions, p 138-167. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch08
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of FIGURE 3
FIGURE 3

Double immunodiffusion profiles of antisera to proteases of . Each well was filled with 10 μl of either an Ag or an antiserum preparation. Wells: A, rabbit antiserum to protease A; B, rabbit antiserum to protease B; C, a 1:1 (vol/vol) mixture of antiserum to protease A and antiserum to protease B, respectively. The Ags were placed into the outer wells. Well 1, protease B from strain 1; well 2, protease A from strain 2; well 3, extracellular products of strain 3. (Courtesy of K. Y. Leung and R. M. W. Stevenson.)

Citation: Mutharia L, Lam J. 2007. Antigen-Antibody Reactions, p 138-167. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch08
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of FIGURE 4
FIGURE 4

XIE of cell lysate and homologous polyclonal antiserum. A 10-μl portion of the lysate was loaded in the first-dimension gel (1D), the intermediate gel (IG) contained NaCl, and the second-dimension gel (2D) contained antiserum.

Citation: Mutharia L, Lam J. 2007. Antigen-Antibody Reactions, p 138-167. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch08
Permissions and Reprints Request Permissions
Download as Powerpoint

References

/content/book/10.1128/9781555817497.chap8
1. Goers, J. 1992. Immunochemical Techniques Laboratory Manual. Academic Press, New York, NY..
2. Hancock, I. C.,, and I. R. Poxton. 1988. Bacterial Cell Surface Techniques. Modern Microbiological Methods. Wiley Interscience, New York, NY. Provides specific protocols for studying immunochemistry of cell surface antigens..
3. Harlow, E. 1999. Using Antibodies: a Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY..
4. Harlow, E.,, and D. Lane. 1988. Antibodies, a Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY..References 3 and 4 are valuable reference books for labs using antibodies..
5. Akita, E. M.,, and S. Nakai. 1992. Immunoglobulin from egg yolk; isolation and purification. J. Food Sci. 57:629634.
6. Herbert, W. J.,, and F. Kristensen,. 1986. Laboratory animal techniques for immunology, p. 133.1133.36. In D. M. Weir,, C. Blackwell,, and L. A. Herzenberg (ed.), Handbook of Experimental Immunology, 4th ed. Blackwell Scientific Publications, Boston, MA..
7. Lee, S. B.,, Y. Mine,, and R. M. W. Stevenson. 2000. Effects of hen egg yolk immunoglobulins in passive protection of rainbow trout against Yersinia ruckeri. J. Agric. Food Chem. 48:110115.
8. Malik, V. S. 1994. Antibody Techniques. Academic Press, New York, NY..
9. Smelser, J. F. 1985. Rabbits: a practical guide for the veterinary technician. Vet. Tech. 6:121128.
10. Stewart, K. L.,, E. L. Johnstone,, and J. A. Vecera. 1988. The laboratory mouse and rat. Vet. Tech. 9:264271.
11. Tillman, P.,, and C. Norman. 1983. Droperidol-fentanyl as an aid to blood collection in rabbits. Lab. Anim. Sci. 33: 181182.
12. Bogard, C. W., Jr.,, D. L. Dunn,, K. Abernthy,, C. Kilgarriff,, and P. C. Kung. 1987. Isolation and characterization of murine monoclonal antibodies specific for gramnegative bacterial lipopolysaccharide: association of crossgenus reactivity with lipid A specificity. Infect. Immun. 55:899908.
13. Bowman, C. C.,, and J. D. Clements. 2001. Differential biological and adjuvant activities of cholera toxin and Escherichia coli heat-labile enterotoxin hybrids. Infect. Immun. 69:15281535.
14. Campbell, D. H.,, J. S. Garvey,, N. E. Cremer,, and D. H. Sussdorf. 1970. Methods in Immunology, 2nd ed. W. A. Benjamin, Inc., New York, NY..
15. Chowdhury, P. S.,, M. Gallo,, and I. Pastan. 2001. Generation of high titre antisera in rabbits by DNA immunization. J. Immunol. Methods 249:147154.
16. Christodoulides, M.,, E. Rattue,, and J. E. Heckles. 2000. Effect of adjuvant on the immune response to a multiple Ag peptide (MAP) containing protective epitopes from Neisseria meningitidis class 1 porin. Vaccine 18:131139.
17. Diano, M.,, A. Le Bivic,, and M. Hirn. 1987. A method for the production of highly specific polyclonal antibodies. Anal. Biochem. 166:224229.
18. Galanos, C.,, O. Luderitz,, and O. Westphal. 1971. Preparation and properties of antisera against the lipid-A component of bacterial lipopolysaccharide. Eur. J. Biochem. 24:116122.
19. Hancock, K.,, and V. C. W. Tsang. 1983. India ink staining of proteins on nitrocellulose paper. Anal. Biochem. 133:157162.
20. Hassan, U. A.,, A. M. Abai,, D. R. Harper,, B. W. Wren,, and W. J. W. Morrow. 1999. Nucleic acid immunization: concepts and techniques associated with third generation vaccines. J. Immunol. Methods 229:122.
21. Hassett, D. E.,, and J. L. Whitton. 1996. DNA immunization. Trends Microbiol. 4:307312.
22. Knudsen, K. A. 1985. Proteins transferred to nitrocellulose for use as immunogen. Anal. Biochem. 147:285288.
23. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680685.
24. Leitner, W. W.,, H. Ying,, and N. P. Restifo. 2000. DNA and RNA-based vaccines: principles, progress and prospects. Vaccine 18:765777.
25. Leppard, K.,, N. Totty,, M. Waterfield,, E. Harlow,, J. Jenkins,, and L. Crawford. 1981. Purification and partial amino acid sequence analysis of the cellular tumor antigen, p53, from mouse SV40-transformed cells. EMBO J. 2:19931999.
26. Lindberg, A. 1999. Glycoprotein conjugate vaccines. Vaccine 17:S28S36.
27. Lipford, G. B.,, K. Heeg,, and H. Wagner. 1998. Bacterial DNA as immune cell activator. Trends Microbiol. 6:496 500.
28. McCluskie, M. J.,, and H. L Davis. 1999. CpG DNA as mucosal adjuvants. Vaccine 18:231237.
29. Rauser, W. E.,, A. A. Quesnel,, J. S. Lam,, and G. G. Southam. 1988. An enzyme-linked immunosorbent assay for plant cadmium binding peptide. Plant Sci. 57:3743.
30. Allison, A. C.,, and G. Gregoriadis. 1974. Liposomes as immunological adjuvants. Nature 252:252.
31. Baker, P. J.,, J. R. Hiernaux,, M. B. Fauntleroy,, B. Prescott,, J. Cantrell,, and J. A. Rubach. 1988. Inactivation of suppressor T-cell activity by nontoxic monophosphoryl lipid A. Infect. Immun. 56:10761083.
32. Baker, P. J.,, J. R. Hiernaux,, M. B. Fauntleroy,, P. W. Stashak,, B. Prescott,, J. L. Cantrell,, and J. A. Tudbach. 1988. Ability of monophosphoryl lipid A to augment the antibody response of young mice. Infect. Immun. 56:30643066.
33. Branche, R.,, and G. Renoux. 1972. Stimulation of rabies vaccine in mice by low doses of polyadenylic: polyuridylic complex. Infect. Immun. 6:324325.
34. Chase, M. W. 1967. Production of antiserum. Methods Immunol. Immunochem. 1:197209.
35. Claassen, E.,, N. Kors,, and N. van Rooijen. 1987. Immunomodulation with liposome: the immune response elicited by liposomes with entrapped dichloromethyldiphosphonate and surface associated antigen or hapten. Immunology 60:509515.
36. De Becker, G.,, V. Moulin,, B. Pajak,, C. Bruck,, M. Francotte,, C. Thiriart,, J. Urbain,, and M. Moser. 2000. The adjuvant monophosphoryl lipid A increases the function of antigen-presenting cells. Int. Immunol. 6:807815.
37. Edelman, R. 1980. Vaccine adjuvants. Rev. Infect Dis. 2:370383.
38. Ellouz, F.,, A. Adam,, R. Ciorbaru,, and E. Lederer. 1974. Minimal structural requirements for adjuvant activity of bacterial peptidoglycan derivatives. Biochem. Biophys. Res. Commun. 59:13171325.
39. Fitzgerald, T. J. 1991. Syphilis vaccine: up-regulation of immunogenicity by cyclophosphamide, Ribi adjuvant, and indomethacin confers significant protection against challenge infection in rabbits. Vaccine 9:266272.
40. Freund, J. 1956. The mode of action of immunological adjuvants. Adv. Tuberc. Res. 7:130148.
41. Gery, I.,, J. Kruger,, and S. Spiesel. 1972. Stimulation of B lymphocytes by endotoxin reactions of thymus-deprived mice and karyotypic analysis of dividing cells in mice bearing T6T6 thymus graft. J. Immunol. 108:10881091.
42. Gregoriadis, G.,, D. Davis,, and A. Davis. 1987. Liposomes as immunological adjuvants: antigen incorporation studies. Vaccine 5:145151.
43. Kenney, J. S.,, B. W. Hughes,, M. P. Masada,, and A. C. Allison. 1989. Influence of adjuvants on the quantity, affinity, isotype and epitope specificity of murine antibodies. J. Immunol. Methods 121:157166.
44. Kripke, M. L.,, and D. W. Weiss. 1970. Studies on the immune responses of Balb/c mice during tumor induction by mineral oil. Intern. J. Cancer 6:422430.
45. Moingeon, P.,, J. Haensler,, and A. Lindberg. 2001. Towards the rational design of Th1 adjuvants. Vaccine 19: 43634372.
46. Morein, B.,, B. Sundquist,, S. Hoglund,, K. Dalsgaard,, and A. Osterhaus. 1984. ISCOM, a novel structure for antigenic presentation of membrane proteins from enveloped viruses. Nature 308:457459.
47. Morein, B.,, K. Lövgren,, S. Höglund,, and B. Sundquist. 1987. The ISCOM: an immunostimulating complex. Immunol. Today 8:333338.
48. Nicholson, K. G.,, D. A. J. Tyrrell,, P. Harrison,, C. W. Potter,, R. Jennings,, A. Clark,, G. C. Scheld,, J. M. Wood,, R. Yelts,, V. Seagroatt,, A. Higgins,, and S. G. Anderson. 1979. Clinical studies of mononvalent inactiviated whole virus and subunit A/USSR/77 (H1 H1) vaccine serological and clinical reactions. J. Biol. Stand. 7:123136.
49. Ohta, M.,, N. Kido,, T. Hasegawa,, H. Ito,, Y. Fujii,, T. Arakawa,, T. Komatsu,, and N. Kato. 1987. Contribution of the mannan O side-chains to the adjuvant action of lipopolysaccharide. Immunology 60:503507.
50. Röonberg, B.,, M. Fekadu,, and B. Morein. 1995. Adjuvant activity of non-toxic Quillaja saponaria Molin components for use in ISCOM matrix. Vaccine 13:13751382.
51. Seppala, I. J. T.,, and O. Makela. 1984. Adjuvant effect of bacterial LPS and/or alum precipitation in response to polysaccharide and protein antigens. Immunology 53:827836.
52. Shek, P. N.,, and B. H. Sabiston. 1981. Immune response mediated by liposome associated protein antigens. I. Potentiation of the plaque forming cell response. Immunology 45:349356.
53. Taub, R. N.,, A. R. Krantz,, and D. W. Dresser. 1970. The effect of localized injection of adjuvant material on the draining lymph node. I. Histology. Immunology 18: 171186.
54. Thoelen, S.,, N. De Clercq,, and N. Tornieporth. 2001. A prophylatic hepatitis B vaccine with a novel adjuvant system. Vaccine 19:24002403.
55. Sjölander, A.,, J. C. Cox,, and I. A. Barr. 1998. ISCOMs: an adjuvant with multiple functions. J. Leukoc. Biol. 64: 713723.
56. Warren, H. S.,, F. R. Vogel,, and L. A. Chedid. 1986. Current status of immunological adjuvants. Annu. Rev. Immunol. 4:369388.
57. Bjorck, L.,, W. Karstern,, G. Lindahl,, and K. Wildeback. 1987. Streptococcal protein G, expressed by streptococci or Escherichia coli, has separate sites for human albumin and IgG. Mol. Immunol. 24:11131122.
58. Bjorck, L.,, and G. Kronvall. 1984. Purification and some properties of streptococcal protein G, a novel IgG-binding reagent. J. Immunol. 133:969974.
59. Bruck, C.,, D. Portetelle,, C. Gilneur,, and A. Bollen. 1982. One step purification of mouse monoclonal antibodies from ascitic fluid by DEAE Affi-gel blue chromatography. J. Immunol. Methods 53:313319.
60. Cassone, A.,, A. Torosantucci,, M. Boccanera,, G. Pellegrini,, C. Palma,, and F. Malavasi. 1988. Production and characterisation of a monoclonal antibody to a cell surface, glucomannoprotein constituent of Candida albicans and other pathogenic Candida species. J. Med. Microbiol. 27:233238.
61. Creuzenet, C.,, and J. S. Lam. 2001. Topological and functional characterisation of WbpM, an inner-membrane UDP-GlcNAc C6 dehydratase essential for lipopolysaccharide biosynthesis in Pseudomonas aeruginosa. Mol. Microbiol. 41:12951310.
62. Davey, M. L.,, R. E. W. Hancock,, and L. M. Mutharia. 1998. Influence of culture conditions on expression of the 40-kilodalton protein of Vibrio anguillarum serotype 02. Appl. Environ. Microbiol. 64:138146.
63. Ey, P. L.,, S. J. Prouse,, and C. R. Jenkin. 1978. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-Sepharose. Immunochemistry 15:429436.
64. Galfré, G.,, and C. Milstein. 1981. Preparation of monoclonal antibodies: strategies and procedure. Methods Enzymol. 73:146.
65. Goding, J. W. 1986. Monoclonal Antibodies: Principle and Practice, 2nd ed. Academic Press, London, England.
66. Köhler, G.,, and C. Milstein. 1975. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495499.
67. Kuhlmann, I.,, W. Kurth,, and I. Ruhdel. 1989. Monoclonal antibodies: in vivo and in vitro production on a laboratory scale, with consideration of the legal aspects of animal protection. Altern. Lab. Anim. 17:7382.
68. Lam, J. S.,, L. A. MacDonald,, M. Y. C. Lam,, L. G. M. Duchesne,, and G. G. Southam. 1987. Production and characterization of monoclonal antibodies against serotype strains of Pseudomonas aeruginosa. Infect. Immun. 55:10511057.
69. Lindmark, R.,, K. Thoren-Tolling,, and J. Sjoquist. 1983. Binding of immunoglobulins to protein A and immunoglobulin levels in mammalian sera. J. Immunol. Methods 62:113.
70. Mariani, M.,, M. Cianfriglia,, and A. Cassone. 1989. Is mouse IgM purification on protein A possible? Immunol. Today 10:115116. (Letter to the editor.)
71. Oi, V. T.,, and L. A. Herzenberg,. 1980. Immunoglobulinproducing hybrid cell lines, p. 351372. In B. B. Mishell, and S. M. Shiigi (ed.), Selected Methods in Cellular Immunology. W. H. Freeman, San Francisco, CA.
72. Tung, A. S.,, S.-T. Ju,, S. Sato,, and A. Nisonoff. 1976. Production of large amounts of antibodies in individual mice. J. Immunol. 116:676681.
73. Wilner, M. A. E.,, H. D. Troutman,, F. W. Trader,, and I. W. McLean. 1963. Vaccine potentiation by emulsification with pure hydrocarbon compounds. J. Immunol. 91: 210229.
74. Edwards, P. R.,, and E. W. H. Ewing. 1962. Identification of Enterobacteriaceae,2nd ed. Burgess Publishing Co., Minneapolis, MN..
75. Fung, J. C.,, and R. C. Tilton,. 1985. Detection of bacterial antigens by counter immunoelectrophoresis, coagglutination and latex agglutination, p. 883890. In E. H. Lennette,, A. Balows,, W. J. Hauser, Jr.,, and W. J. Shadomy (ed.), Manual of Clinical Microbiology, 4th ed. American Society for Microbiology, Washington, DC..
76. Handfield, S. G.,, A. Lane,, and M. B. McIllmurray. 1987. A novel coloured latex test for detection and identification of more than one antigen. J. Immunol. Methods 97: 153158.
77. Kronvall, G. 1973. A rapid slide agglutination method for typing pneumococci by means of specific antibody adsorbed to protein A-containing staphylococci. J. Med. Microbiol. 6:187190.
78. Lim, P. L.,, and K. H. Ko. 1990. A tube latex test based on colour separation for detection of IgM antibodies to either of two different microorganisms. J. Immunol. Methods 135:914.
79. McIllmurray, M. B.,, and M. D. Moody,. 1986. Latex agglutination, p. 928. In R. B. Kohler (ed.), Antigen Detection To Diagnose Bacterial Infections, vol 1. CRC Press, Boca Raton, FL..
80. Oakley, C. L., 1971. Antigen-antibody reactions in microbiology, p. 174217. In J. R. Norris, and D. W. Ribbons (ed.), Methods in Microbiology. Academic Press, London, England.
81. Merill, C. W.,, J. M. Gwaltney,, J. O. Hendley,, and M. A. Sande. 1973. Rapid identification of pneumococci. N. Engl. J. Med. 288:510512.
82. Bjerrum, O. J.,, and T. C. Bøg-Hansen. 1975. Immunochemical gel precipitation techniques in membrane studies p. 378426. In A. H. Maddy (ed.), Biochemical Analysis of Membranes. John Wiley and Sons, Inc., New York, NY..
83. Laurell, C. B. 1966. Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal. Biochem. 15:4552.
84.Ouchterlony, Ö. 1968. Handbook of Immunodiffusion and Immunoelectrophoresis. Ann Arbor Science Publishers, Ann Arbor, MI.
85. Tilton, R., 1983. Procedures for the detection of microorganisms by counter immunoelectrophoresis, p. 8796. In J. D. Coonrod,, L. J. Kunz,, and M. J. Ferraro (ed.), The Direct Detection of Microorganisms in Clinical Samples. Academic Press, Orlando, FL.
86. Weeke, B., 1973. Crossed immunoelectrophoresis, p. 4556. In N. H. Axelsen,, J. Krøll,, and B. Weeke (ed.), A Manual of Quantitative Immunoelectrophoresis. Blackwell Scientific Publications, Oxford, England.
87. Cantarero, L. A.,, J. E. Butler,, and J. W. Osborne. 1980. The adsorptive characteristic of proteins for polystyrene and their significance in solid phase immunoassays. Anal. Biochem. 105:375382.
88. Conradie, J. D.,, M. Govender,, and L. Visser. 1983. ELISA solid phase: partial denaturation of coating antibody yields a more efficient solid phase. J. Immunol. Methods 59:289299.
89. Engvall, E.,, and P. Perlmann. 1971. Enzyme-linked immunosorbent assay (ELISA): quantitative assay of immunoglobulin. Immunochemistry 8:871879.
90. Fischer, P. M.,, and M. E. H. Howden. 1990. Direct, enzyme- linked immunosorbent assay of anti-peptide antibodies using capture of biotinylated peptides by immobilised avidin. J. Immunoassay 11:311327.
91. Gardas, A.,, and A. Lewartowska. 1988. Coating of proteins to polystyrene ELISA plates in the presence of detergents. J. Immunol. Methods 106:251255.
92. Ito, J. I., Jr.,, A. C. Wunderlich,, J. Lyons,, C. E. Davis,, D. G. Gurney,, and A. I. Braude. 1980. Role of magnesium in the enzyme-linked immunosorbent assay for lipopolysaccharides of rough Escherichia coli strain J5 and Neisseria gonorrhoeae. J. Infect. Dis. 142:532537.
93. Julian, E.,, M. Cama,, P. Martínez,, and M. Luquin. 2001. An ELISA for five glycolipids form the cell wall of Mycobacterium tuberculosis: Tween 20 interference in the assay. J. Immunol. Methods 251:2130.
94. Lee, B. Y.,, D. Chatterjee,, C. M. Bozic,, P. J. Brennan,, D. L. Cohn,, J. D. Bales,, S. M. Harrison,, L. A. Androu,, and I. M. Orme. 1991. Prevalence of serum antibody to the type-specific glycopeptidolipid antigens of Mycobacterium avium in human immunodeficiency viruspositive and -negative individuals. J. Clin. Microbiol. 29: 10261029.
95. Lewkowich, I. P.,, J. D. Campbell,, and K. T. HayGlass. 2001. Comparison of chemiluminescent assays and colorimetric ELISAs for quantification of murine IL-12, human IL-4 and murine IL-4: chemiluminescent substrates provide markedly enhanced sensitivity. J. Immunol. Methods 247:111118.
96. Lutz, H. U.,, P. Stammler,, and E. A. Fischer. 1990. Covalent binding of detergent-solubilised membrane glycoproteins to “Chemobond” plates for ELISA. J. Immunol Methods 129:211220.
97. Munoz, C.,, A. Nieto,, A. Gaya,, J. Martinez,, and J. Vives. 1986. New experimental criteria for optimisation of solid phase antigen concentration and stability in ELISA. J. Immunol. Methods 94:137144.
98. Nakamura, R. M.,, A. Valler,, and D. E. Bidwell,. 1986. Enzyme immunoassays: heterogeneous and homogenous systems, p. 27.127.2. In D. M. Weir,, C. Blackwell,, and L. A. Herzenberg (ed.), Handbook of Experimental Immunology, 4th ed., vol. 1. Immunochemistry. Blackwell Scientific Publications, Boston, MA.
99. Pitzurra, L.,, E. Blast,, A. Bartoli,, P. Marconi,, and F. Bistoni. 1990. A rapid objective immunofluorescence microassay. Application for detection of surface and intracellular antigens. J. Immunol. Methods 135:7175.
100. Reggiardo, Z.,, E. Vasquez,, and L. Schnaper. 1980. ELISA tests for antibodies against mycobacterial glycolipids. J. Immunol. Methods 34:5560.
101. Rongen, H. A.,, H. M. van der Horst,, A. J. van Oosterhout,, A. Bult,, and W. P. van Bennekom. 1996. Application of xanthine oxidase-catalysed luminol chemiluminescence in a mouse interleukin-5 immunoassay. J. Immunol. Methods 197:161169.
102. Scott, B. B.,, and G. R. Barclay. 1987. Endotoxinpolymyxin complexes in an improved enzyme-linked immunosorbent assay for IgG antibodies in blood donor sera to gram-negative endotoxin core glycolipids. Vox Sang. 52:272280.
103. Tijseen, P., 1985. Chapter 1, p. 9327. In R. H. Burdon, and P. H. Van Knippenberg (ed.), Laboratory Techniques in Biochemistry and Molecular Biology. Practice and Theory of Enzyme Immunoassays. Elsevier Science Publishing Company, Inc., New York, NY..
104. Voller, A.,, and D. E. Bidwell,. 1985. Enzyme immunoassays, p. 7786. In W. P. Collins (ed.), Alternate Immunoassays. John Wiley & Sons, Inc., New York, NY..
105. Weigand, K.,, C. Birr,, and M. Suter. 1981. The hexaand pentapeptide extension of proalbumin. II. Processing of specific antibodies against the synthetic hexapeptide. Biochim. Biophys. Acta 670:424427.
106. Wylie, D. E.,, L. D. Carlson,, R. Carlson,, F. W. Wagner,, and S. M. Schuster. 1991. Detection of mercuric ions in water by ELISA with a mercury-specific antibody. Anal. Biochem. 194:381387.
107. Zhao, X.,, and J. S. Lam. 2001. WaaP of Pseudomonas aeruginosa is a novel eukaryotic type protein-tyrosine kinase as well as a sugar kinase essential for the biosynthesis of core lipopolysaccharide. J. Biol. Chem. 277:47224730.
108. Birk, H. W.,, and H. Koepsell. 1987. Reaction of monoclonal antibodies with plasma membrane proteins after binding to nitrocellulose: renaturation of antigenic sites and reduction of non-specific antibody binding. Anal. Biochem. 164:1222.
109. Houston, B.,, and D. Peddie. 1989. A method for detecting proteins immobilised on nitrocellulose membranes by in situ derivatization with fluorescein isothiocyanate. Anal. Biochem. 177:263267.
110. Mutharia, L. M.,, and M. Steele. 1995. Characterization of concanavalin A-binding glycoproteins from procyclic culture forms of Trypanosoma congolense, T. simiae and T. brucei brucei. Parasitol Res. 81:245252.
111. O’Shannessy, D. J.,, P. J. Voorstad,, and R. H. Quarles. 1987. Quantitation of glycoproteins on electroblots using the biotin-streptavidin complex. Anal. Biochem. 163: 204209.
112. Schaffner, W.,, and C. Weissman. 1973. A rapid, sensitive and specific method for the determination of protein in dilute solution. Anal. Biochem. 56:502514.
113. Towbin, M.,, T. Staehlin,, and J. Gordon. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some application. Proc. Natl. Acad. Sci. USA 76:43504354.
114. Woodward, M. P.,, W. W. Young, Jr.,, and R. A. Bloodgood. 1985. Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation. J. Immunol. Methods 78:143153.
115. Herzenberg, L. A.,, S. C. De Rosa,, and L. A. Herzenberg. 2000. Monoclonal antibodies and the FACS: complementary tools for immunobiology and medicine. Immunol. Today 21:383390.
116. Johnson, G. D.,, and E. J. Holborow,. 1986. Preparation and use of fluorochrome conjugates, p. 18.118.20. In D. M. Weir,, C. Blackwell,, and L. A. Herzenberg (ed.), Handbook of Experimental Immunology, 4th ed., vol. 1. Immunochemistry. Blackwell Scientific Publications, Boston, MA..
117. McKinney, R. M., 1986. Immunofluorescence microscopy: reagents and technique, p. 3549. In R. B. Kohler (ed.), Antigen Detection To Diagnose Bacterial Infections, vol. 1. Methods. CRC Press, Boca Raton, FL.

Tables

Generic image for table
TABLE 1

Volumes of Ag-Ab mixtures for injection

Freund's adjuvants should not be used with i.v. injections.

Citation: Mutharia L, Lam J. 2007. Antigen-Antibody Reactions, p 138-167. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch08
Generic image for table
TABLE 2

Adjuvants and their characteristics

Citation: Mutharia L, Lam J. 2007. Antigen-Antibody Reactions, p 138-167. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch08
Generic image for table
TABLE 3

Commercial sources of immunoglobulin isotype determination reagents

Citation: Mutharia L, Lam J. 2007. Antigen-Antibody Reactions, p 138-167. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch08
Generic image for table
TABLE 4

Conditions for HPLC purification of various immunoglobulin isotypes

Citation: Mutharia L, Lam J. 2007. Antigen-Antibody Reactions, p 138-167. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch08

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error