Purification of monoclonal antibodies using high performance liquid chromatography (HPLC)

G. A. Perry, J. D. Jackson, Thomas L McDonald, D. A. Crouse, J. G. Sharp

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Recent increases in the ability to detect low levels of immunofluorescence have shown the need for highly purified primary immunoreagents. There are now reports of purification of monoclonal antibodies using HPLC with reverse phase columns. In this study we have utilized standard size exclusion HPLC to purify both biotinylated and non-biotinylated monoclonal antibodies from hybridoma culture supernatants. Results indicated that both biotinylated and non-biotinylated monoclonal antibodies retained their antigen binding capacity after purification, and were not different in this capacity from commercially available, affinity purified reagents. These findings indicate that size exclusion HPLC may be used in the purification of biologically active monoclonal antibodies, and suggest that this technique may be used in the large scale production of antibodies and their fragments, in antibody purification from ascites fluid, and in antisera quality control.

Original languageEnglish (US)
Pages (from-to)431-447
Number of pages17
JournalPreparative Biochemistry
Volume14
Issue number5
DOIs
StatePublished - Dec 1 1984

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High performance liquid chromatography
Purification
High Pressure Liquid Chromatography
Monoclonal Antibodies
Immunoglobulin Fragments
Hybridomas
Ascites
Quality Control
Fluorescent Antibody Technique
Quality control
Immune Sera
Antigens
Fluids
Antibodies

ASJC Scopus subject areas

  • Biochemistry
  • Genetics

Cite this

Purification of monoclonal antibodies using high performance liquid chromatography (HPLC). / Perry, G. A.; Jackson, J. D.; McDonald, Thomas L; Crouse, D. A.; Sharp, J. G.

In: Preparative Biochemistry, Vol. 14, No. 5, 01.12.1984, p. 431-447.

Research output: Contribution to journalArticle

Perry, G. A. ; Jackson, J. D. ; McDonald, Thomas L ; Crouse, D. A. ; Sharp, J. G. / Purification of monoclonal antibodies using high performance liquid chromatography (HPLC). In: Preparative Biochemistry. 1984 ; Vol. 14, No. 5. pp. 431-447.
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