Characterization of antibody covalently coupled to liposomes

Anthony Huang, Y. S. Tsao, Stephen Kennel, Leaf Huang

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

We have explored the covalent coupling of fatty acids to immunoglobulin G(IgG). N-hydroxysuccinimide ester of palmitic acid (NHSP) was used to couple palmitic acid to either a mouse monoclonal antibody to the major histocompatibility antigen, H-2 k , or goat antibody to the major glycoprotein of the Molony Leukemia Virus, gp-70. The reaction was characterized in terms of the time course, input ratio of NHSP to IgG, stoichiometry of the coupling, distribution of palmitic acid in the IgG subunits, and the antigen binding capacity of the coupled antibody. Incorporation of the fatty acid modified IgG into liposomal membranes using a detergent-dialysis method was studied as a function of extent of fatty acid coupling. Finally, the binding of IgG-coated liposomes with cells expressing proper antigens was characterized. The major conclusions were: (1) the optimal molar ratio of NHSP to IgG in the reaction was between 10 and 20, which yields about 4-5 palmitoyl chains per IgG molecule; (2) at this level of coupling, the antigen binding capacity of the IgG antibody decreased about 3-4-fold; (3) incorporation of the coupled antibody into unilamellar liposomes (about 1000 Å diameter) can be achieved with a deoxycholate-dialysis method with an optimal lipid-to-protein ratio of 10:1 (w/w); (4) there were about 48 IgG molecules incorporated per liposome under these conditions; (5) the apparent dissociation constant of the liposome-bound antibody under the optimal condition was about 6-7-fold higher than that of the native antibody; (6) binding of antibody to the target cells was accompanied by binding of liposomal lipids; both bindings could be blocked by pretreatment of cell with unmodified antibody.

Original languageEnglish (US)
Pages (from-to)140-150
Number of pages11
JournalBBA - General Subjects
Volume716
Issue number2
DOIs
StatePublished - May 27 1982
Externally publishedYes

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Liposomes
Immunoglobulin G
Palmitic Acid
Antibodies
Esters
Fatty Acids
Dialysis
Antigens
Immunoglobulin Subunits
Cells
Lipids
Unilamellar Liposomes
Histocompatibility Antigens
Molecules
Deoxycholic Acid
Viruses
Goats
Stoichiometry
Detergents
Glycoproteins

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Characterization of antibody covalently coupled to liposomes. / Huang, Anthony; Tsao, Y. S.; Kennel, Stephen; Huang, Leaf.

In: BBA - General Subjects, Vol. 716, No. 2, 27.05.1982, p. 140-150.

Research output: Contribution to journalArticle

Huang, Anthony ; Tsao, Y. S. ; Kennel, Stephen ; Huang, Leaf. / Characterization of antibody covalently coupled to liposomes. In: BBA - General Subjects. 1982 ; Vol. 716, No. 2. pp. 140-150.
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