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High-performance liquid affinity chromatography: rapid immunoanalysis of transferrin in serum

Ohlson, Sten ; Gudmundsson, Britt-Marie ; Wikström, Per and Larsson, Per-Olof LU (1988) In Clinical Chemistry 34(10). p.2039-2043
Abstract
We describe a new method for quantitatively measuring substances of
clinical interest by high-performance liquid affinity chromatography
(HPLAC). As a model system we selected analysis for transferrin in human
serum with immobilized antibodies in a high-performance liquid
chromatographic system. SelectiSpher-10 Activated Tresyl columns (5 or
10 x 0.5 cm) were used for in situ coupling of polyclonal antibodies to
transferrin. The amount of transferrin eluted was determined by
integrating the eluted peak at 280 nm. The whole analytical
procedure--including injection of sample, washing, elution, and analysis
of data--takes only 7 min. We characterized the HPLAC system for
analysis of... (More)
We describe a new method for quantitatively measuring substances of
clinical interest by high-performance liquid affinity chromatography
(HPLAC). As a model system we selected analysis for transferrin in human
serum with immobilized antibodies in a high-performance liquid
chromatographic system. SelectiSpher-10 Activated Tresyl columns (5 or
10 x 0.5 cm) were used for in situ coupling of polyclonal antibodies to
transferrin. The amount of transferrin eluted was determined by
integrating the eluted peak at 280 nm. The whole analytical
procedure--including injection of sample, washing, elution, and analysis
of data--takes only 7 min. We characterized the HPLAC system for
analysis of transferrin in several ways: intra-assay CV approximately
3%; inter-assay CV 2-9%; linear response up to 1 mg/mL column volume;
detection limit approximately 3 micrograms; analytical recovery 98% +/-
2%; purity of eluted sample greater than 95% (SDS-PAGE). The HPLAC
method was compared with "rocket" immunoelectrophoresis, a commonly used
method of analysis for transferrin, and there was excellent correlation
between the two methods (r = 0.96, n = 60). Benefits of this HPLAC
technique include high precision, rapid analysis, and simplified sample
handling. (Less)
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author
; ; and
publishing date
type
Contribution to journal
publication status
published
subject
keywords
HPLAC, Transferrin, Clinical chemistry
in
Clinical Chemistry
volume
34
issue
10
pages
5 pages
publisher
Oxford University Press
external identifiers
  • scopus:0023711899
ISSN
0009-9147
DOI
10.1093/clinchem/34.10.2039
language
English
LU publication?
no
id
eb599f13-8ff9-4ef3-9d2b-0f940ab47dfa
date added to LUP
2024-06-18 13:43:33
date last changed
2025-04-04 14:28:08
@article{eb599f13-8ff9-4ef3-9d2b-0f940ab47dfa,
  abstract     = {{We describe a new method for quantitatively measuring substances of <br>
clinical interest by high-performance liquid affinity chromatography <br>
(HPLAC). As a model system we selected analysis for transferrin in human<br>
 serum with immobilized antibodies in a high-performance liquid <br>
chromatographic system. SelectiSpher-10 Activated Tresyl columns (5 or <br>
10 x 0.5 cm) were used for in situ coupling of polyclonal antibodies to <br>
transferrin. The amount of transferrin eluted was determined by <br>
integrating the eluted peak at 280 nm. The whole analytical <br>
procedure--including injection of sample, washing, elution, and analysis<br>
 of data--takes only 7 min. We characterized the HPLAC system for <br>
analysis of transferrin in several ways: intra-assay CV approximately <br>
3%; inter-assay CV 2-9%; linear response up to 1 mg/mL column volume; <br>
detection limit approximately 3 micrograms; analytical recovery 98% +/- <br>
2%; purity of eluted sample greater than 95% (SDS-PAGE). The HPLAC <br>
method was compared with "rocket" immunoelectrophoresis, a commonly used<br>
 method of analysis for transferrin, and there was excellent correlation<br>
 between the two methods (r = 0.96, n = 60). Benefits of this HPLAC <br>
technique include high precision, rapid analysis, and simplified sample <br>
handling.}},
  author       = {{Ohlson, Sten and Gudmundsson, Britt-Marie and Wikström, Per and Larsson, Per-Olof}},
  issn         = {{0009-9147}},
  keywords     = {{HPLAC; Transferrin; Clinical chemistry}},
  language     = {{eng}},
  number       = {{10}},
  pages        = {{2039--2043}},
  publisher    = {{Oxford University Press}},
  series       = {{Clinical Chemistry}},
  title        = {{High-performance liquid affinity chromatography: rapid immunoanalysis of transferrin in serum}},
  url          = {{http://dx.doi.org/10.1093/clinchem/34.10.2039}},
  doi          = {{10.1093/clinchem/34.10.2039}},
  volume       = {{34}},
  year         = {{1988}},
}