Immunochemical property of human haptoglobin phenotypes: Determination of plasma haptoglobin using type-matched standards

Tsai Mu Cheng, Ju Pin Pan, Shiau Ting Lai, Li Pin Kao, Hong Huei Lin, Simon J T Mao

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Objectives: Haptoglobin (Hp) phenotypes 1-1, 2-1, and 2-2 are associated with inflammatory diseases. Since their biochemical structures are rather heterogeneous, it is necessary to accurately determine the plasma Hp levels. Design and methods: Immunodiffusion, immunoturbidimetric, and noncompetitive ELISA were conducted to determine the differences in immunoreactivity among Hp phenotypes and to verify that such difference may significantly affect the outcome of Hp determinations. A novel ELISA using phenotype-matched calibrators was performed to compared with a commercial GenWay ELISA kit using a single calibrator in normal healthy males. Results: In immunodiffusion and immunoturbidimetric assays, the immunoreactivity of Hp 1-1 was markedly higher than 2-1 and 2-2, while an opposite result was observed using an ELISA. The latter was primarily due to the repeated antigenic epitopes in polymeric 2-1 and 2-2. Thus, Hp levels could be significantly over- or underestimated depending on the method. An accurate ELISA could be achieved when using each type-specific Hp calibrator matched to each type subject. We show the mean levels of Hp 1-1 subjects (n = 16; 184 ± 42 mg/dL) to be significantly and differentially greater than 2-1 (n = 28; 153 ± 55 mg/dL) (p <0.05) and 2-2 (n = 24; 93 ± 54 mg/dL) (p <0.01) subjects. Conclusions: Due to the diverse immunochemical structure among the Hp types, phenotyping should be performed in all the patients and a type-matched Hp calibrator should be used in clinical Hp determination.

Original languageEnglish
Pages (from-to)1045-1056
Number of pages12
JournalClinical Biochemistry
Volume40
Issue number13-14
DOIs
Publication statusPublished - Sep 2007
Externally publishedYes

Fingerprint

Haptoglobins
Phenotype
Plasmas
Enzyme-Linked Immunosorbent Assay
Immunodiffusion
human HP protein
Epitopes
Assays

Keywords

  • αβ and αβ chain expression
  • ELISA
  • Haptoglobin phenotypes
  • HepG2
  • Immunoreactivity
  • Plasma concentration

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Immunochemical property of human haptoglobin phenotypes : Determination of plasma haptoglobin using type-matched standards. / Cheng, Tsai Mu; Pan, Ju Pin; Lai, Shiau Ting; Kao, Li Pin; Lin, Hong Huei; Mao, Simon J T.

In: Clinical Biochemistry, Vol. 40, No. 13-14, 09.2007, p. 1045-1056.

Research output: Contribution to journalArticle

Cheng, Tsai Mu ; Pan, Ju Pin ; Lai, Shiau Ting ; Kao, Li Pin ; Lin, Hong Huei ; Mao, Simon J T. / Immunochemical property of human haptoglobin phenotypes : Determination of plasma haptoglobin using type-matched standards. In: Clinical Biochemistry. 2007 ; Vol. 40, No. 13-14. pp. 1045-1056.
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abstract = "Objectives: Haptoglobin (Hp) phenotypes 1-1, 2-1, and 2-2 are associated with inflammatory diseases. Since their biochemical structures are rather heterogeneous, it is necessary to accurately determine the plasma Hp levels. Design and methods: Immunodiffusion, immunoturbidimetric, and noncompetitive ELISA were conducted to determine the differences in immunoreactivity among Hp phenotypes and to verify that such difference may significantly affect the outcome of Hp determinations. A novel ELISA using phenotype-matched calibrators was performed to compared with a commercial GenWay ELISA kit using a single calibrator in normal healthy males. Results: In immunodiffusion and immunoturbidimetric assays, the immunoreactivity of Hp 1-1 was markedly higher than 2-1 and 2-2, while an opposite result was observed using an ELISA. The latter was primarily due to the repeated antigenic epitopes in polymeric 2-1 and 2-2. Thus, Hp levels could be significantly over- or underestimated depending on the method. An accurate ELISA could be achieved when using each type-specific Hp calibrator matched to each type subject. We show the mean levels of Hp 1-1 subjects (n = 16; 184 ± 42 mg/dL) to be significantly and differentially greater than 2-1 (n = 28; 153 ± 55 mg/dL) (p <0.05) and 2-2 (n = 24; 93 ± 54 mg/dL) (p <0.01) subjects. Conclusions: Due to the diverse immunochemical structure among the Hp types, phenotyping should be performed in all the patients and a type-matched Hp calibrator should be used in clinical Hp determination.",
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