High-performance liquid affinity chromatography: rapid immunoanalysis of transferrin in serum
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/eb599f13-8ff9-4ef3-9d2b-0f940ab47dfa
- author
- Ohlson, Sten ; Gudmundsson, Britt-Marie ; Wikström, Per and Larsson, Per-Olof LU
- publishing date
- 1988
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- HPLAC, Transferrin, Clinical chemistry
- in
- Clinical Chemistry
- volume
- 34
- issue
- 10
- pages
- 5 pages
- publisher
- American Association for Clinical Chemistry
- 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
- 2024-09-04 09:37:41
@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 = {{American Association for Clinical Chemistry}}, 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}}, }