Detection of serum haptoglobin by enzyme-linked immunosorbent assay in cows with fatty liver

Nakagawa, H; Yamamoto, Osamu; Oikawa, Shin; Higuchi, Hidetoshi; Watanabe, Atsushi; Katoh, Norio

doi:10.1016/s0034-5288(97)90135-1

Cited by 35 publications

(38 citation statements)

References 22 publications

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“…However, this method showed great variability [25] and, moreover, was influenced by the concentration of serum albumin [14]. The ELISA used in the present study was sensitive and was not influenced by the concentration of serum albumin.…”

Section: Discussionmentioning

confidence: 69%

“…Moreover, human Hp subtype patterns significantly correlate with serum lipid concentrations [2]. In cattle, Hp is detected in serum from cows with fatty liver [14,23], and is induced by fatty liver-producing agents, ethionine [18] and glucocorticoids [5,24]. We have recently shown that Hp [8] and Hb [8,21] are distributed in the lipoprotein fractions, particularly in HDL and very high-density lipoprotein (VHDL) fractions, and concentration ratios of Hp in the lipoprotein fractions are 0.2 to 0.3% of total serum Hp, as indicated by densitometric scanning of immunoblotting.…”

mentioning

confidence: 99%

“…The serum Hp concentration has been determined by single radial immunodiffusion (SRID) [12,24] and enzymelinked immunosorbent assay (ELISA) [14,17]. SRID is too insensitive to determine the Hp concentrations in the lipoprotein fractions.…”

mentioning

confidence: 99%

“…SRID is too insensitive to determine the Hp concentrations in the lipoprotein fractions. The ELISA, in which test serum is directly coated on microplates, is sensitive, but its reliability is influenced by the serum albumin concentration [14]. An ELISA based on affinity with Hb has been developed [25].…”

mentioning

confidence: 99%

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Analysis by Enzyme-Linked Immunosorbent Assay and 2-Dimensional Electrophoresis of Haptoglobin in the High-Density Lipoprotein Fraction in Cows.

Kanno

Katoh

2001

The Journal of Veterinary Medical Science

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ABSTRACT. Haptoglobin (Hp) is a hemoglobin (Hb)-binding acute-phase protein. Besides its relevance in inflammation, Hp is involved in the regulation of lipid metabolism. In cattle, in addition to the lipoprotein-deficient fraction, Hp is distributed in high-density lipoprotein (HDL) and very high-density lipoprotein (VHDL) fractions. The purpose of this study was to determine Hp concentrations in the lipoprotein fractions using an enzyme-linked immunosorbent assay (ELISA) based on the affinity with Hb, and also to detect structural differences of HDL Hp from that in the lipoprotein-deficient fraction using 2-dimensional electrophoresis. When purified Hp was used as the antigen for the ELISA, the detection limit was 7.4 ng/ml and linearity was obtained from 14.8 to 475 ng/ml. The correlation coefficient between the ELISA and single radial immunodiffusion was 0.884. The ELISA was shown to be applicable to evaluate Hp concentrations in the lipoprotein fractions. Hp concentrations in the lipoprotein fractions were in the range of 0.94 to 8.77 µg of Hp/ml (n=4), and concentration ratios were 0.2 to 0.3% of whole serum Hp. Of the lipoprotein fractions, Hp was most abundant in HDL, moderate in VHDL and faint in chylomicrons, the very low-density lipoprotein fraction and low-density lipoprotein fraction. By 2-dimensional electrophoresis, α-and β-chains of serum Hp were each separated into 5 spots, and their isoelectric point (pI) values were from 5.05 to 6.28 in the α-chain and from 5.92 to 6.95 in the β-chain. The pI values of HDL Hp were indistinguishable from those of serum Hp. These results indicate that the ELISA based on the affinity with Hb is useful for evaluating Hp concentrations in lipoprotein fractions, and also suggest that HDL Hp is structurally similar to that in the lipoprotein-deficient fraction. KEY WORDS: bovine, enzyme-linked immunosorbent assay, haptoglobin, high-density lipoprotein, 2-dimensional electrophoresis.

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Section: Discussionmentioning

confidence: 69%

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confidence: 99%

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confidence: 99%

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confidence: 99%

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Analysis by Enzyme-Linked Immunosorbent Assay and 2-Dimensional Electrophoresis of Haptoglobin in the High-Density Lipoprotein Fraction in Cows.

Kanno

Katoh

2001

The Journal of Veterinary Medical Science

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“…The elevation of serum concentrations of acute phase proteins by stimuli other than inflammation is not restricted to apoSAA. The serum concentration of haptoglobin, another major acute-phase protein in bovines, is increased in cattle subjected to transportation exhaustion [11,13], in cows with fatty liver [14,20,22,27], in cows to which dexamethasone is administered [6,28], in cows during nonfeeding [28] and in feedlot cattle [29]. The acute phase proteins are induced in response to a wide variety of stimuli, and the response does not distinguish the kind of stimulus.…”

Section: Discussionmentioning

confidence: 99%

The Presence of Two Low Molecular Mass Proteins Immunologically Related to 14 Kilodalton Serum Amyloid A in the Lipoprotein Fraction and Their Decreased Serum Concentrations in Calves with Experimentally Induced Pneumonia

Yamamoto

Katoh

Adachi

1998

The Journal of Veterinary Medical Science

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ABSTRACT. A 14 kilodalton (kDa) serum amyloid A (apoSAA) protein was purified from cow serum. Rabbit antiserum to the 14 kDa apoSAA recognized, in addition to the 14 kDa protein, a 7.5-9.0 kDa protein and a protein having a molecular mass of less than 6.5 kDa (<6.5 kDa protein). The possibility that the two proteins were contaminants was excluded by results showing that the two proteins detected in early stages of purification procedures were not found in the purified 14 kDa apoSAA fraction, as revealed by immunoblot analysis. As in the 14 kDa apoSAA, the 7.5-9.0 kDa protein was localized in the high-density lipoprotein fraction, while the <6.5 kDa protein was in the low-density lipoprotein fraction. In calves with pneumonia induced by inoculation to the lungs of Pasteurella haemolitica, the serum concentration of the 14 kDa apoSAA was increased, whereas those of the 7.5-9.0 kDa and the <6.5 kDa proteins were conversely decreased. The time-course study indicated that the increase in concentration of the 14 kDa apoSAA and decrease in that of the <6.5 kDa protein occurred almost simultaneously. These results suggest that the 14 kDa apoSAA and the immunologically related 7.5-9.0 kDa and <6.5 kDa proteins act as positive and negative acute phase reactants, respectively, and also that concentrations of the three proteins are regulated in concert in acute phase plasma. fraction and, moreover, the serum concentration of the 14 kDa apoSAA was increased whereas those of the two low molecular mass proteins were conversely decreased in sera from calves with pneumonia induced by inoculation of Pasteurella haemolitica. MATERIALS AND METHODS Purification of apoSAA:ApoSAA was purified from cow serum during the course of purification of apoC-III [26]. Briefly, the total lipoprotein fraction [d<1.21, containing chylomicrons (CM) to HDL] prepared from sera of Holstein cows during midlactation was treated with an equal volume of acetone to extract low molecular mass apolipoproteins such as apoC-III and apoSAA. The supernatant obtained by centrifugation at 140 × g for 10 min was lyophilized and thereafter delipidated by mixing with 20 volumes of 2-propanol. Apolipoproteins pelleted by centrifugation at 1,300 × g for 10 min, were dissolved in 6 M urea and 50 mM Tris-HCl (pH 8.6), and applied to a column (2.5 × 90 cm) of Sephadex G-75 (Pharmacia Biotech, Uppsala, Sweden). Fractions rich in low molecular mass apolipoproteins were next applied to a DEAE-Sephacel (Pharmacia Biotech, Uppsala, Sweden) column (2 × 25 cm), and bound proteins were eluted by use of a linear gradient of NaCl (0-0.125 M, total 1 liter). Fractions containing apoSAA were dialyzed against 10 mM ammonium hydrogen carbonate, using cellulose dialyzer tubing with a 3,500 molecular weight cutoff, after which the dialysate was

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Potential serum biomarkers associated with mild and severe leptospirosis infection: A cohort study in the Malaysian population

et al. 2017

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Leptospirosis is an emerging zoonotic infectious disease in Malaysia. The symptoms of leptospirosis vary from mild nonspecific flu-like illness to a severe condition which is usually associated with serious complication and fatality. To study the protein expression profile of mild and severe leptospirosis, 15 paired sera were collected from the patients who were mildly infected and following that progressed to severe stage. The proteome profiles of mild and severe cases were studied using 2DE analysis in combination with LC-MS/MS. The expression of proteins that were significantly different and had a fold difference of at least 2 had been identified and then validated using Western blot. Our study demonstrated apolipoprotein A-I (APOA-I), serum amyloid A (SAA), transferrin (TF), haptoglobin (HP) and transthyretin (TTR) have significantly different expression between mild and severe leptospirosis. The Ingenuity Pathway Analysis software suggested the expression of these five proteins were modulated by acute phase response signaling pathway. Besides that, a functional network of lipid metabolism, molecular transport and small molecule biochemistry that interconnects these five proteins with interactomes also had been predicted by this software. In conclusion, this finding supports the potential of these five proteins to be the biomarkers for mild and severe human leptospirosis.

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Detection of serum haptoglobin by enzyme-linked immunosorbent assay in cows with fatty liver

Cited by 35 publications

References 22 publications

Analysis by Enzyme-Linked Immunosorbent Assay and 2-Dimensional Electrophoresis of Haptoglobin in the High-Density Lipoprotein Fraction in Cows.

Analysis by Enzyme-Linked Immunosorbent Assay and 2-Dimensional Electrophoresis of Haptoglobin in the High-Density Lipoprotein Fraction in Cows.

The Presence of Two Low Molecular Mass Proteins Immunologically Related to 14 Kilodalton Serum Amyloid A in the Lipoprotein Fraction and Their Decreased Serum Concentrations in Calves with Experimentally Induced Pneumonia

Potential serum biomarkers associated with mild and severe leptospirosis infection: A cohort study in the Malaysian population

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