α-1 Antitrypsin phenotypes determined by isoelectric focusing of the cysteine-antitrypsin mixed disulfide in serum

Pierce, John A.; Jeppsson, J.-O.; Laurell, C.‐B.

doi:10.1016/0003-2697(76)90327-4

Cited by 67 publications

(14 citation statements)

References 17 publications

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“…For some of these subjects duplicate samples were also sent to a reference laboratory for corroboration of our results. To obtain an enriched and partially purified a 1 AT from human serum, some modifications were introduced to a method previously described [26] in order to simplify and automate the procedure. In synthesis, serum samples (800 mL to 1 mL) were mixed with 100 mL of 0.02 M DTT in acetate buffer (0.1 M, pH 6.0) and, after 5 min, the mixture was applied to a column (2 cm´1.5 cm ID; total bed volume, 3.75 mL) containing Activated Thiol Sepharose 4B from Pharmacia (Uppsala, Sweden).…”

Section: Sample Preparationmentioning

confidence: 99%

α1-Antitrypsin in serum determined by capillary isoelectric focusing

et al. 2000

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A capillary isoelectric focusing (CIEF) method using bare fused-silica capillaries filled with polyethylene oxide (PEO) and carrier ampholyte solutions in the pH 3.5-5.0 range has been developed for the identification of alpha1-antitrypsin (alpha1AT) phenotypes in human serum. This novel procedure was routinely applied to the study of serum samples of five controls whose alpha1AT phenotype was previously identified and of twelve subjects whose alpha1AT phenotype was unknown. The results obtained allowed us to confirm or identify the alpha1AT phenotype in all sera tested. This procedure seems particularly suitable for identification of alpha1AT variants associated with diseases of clinical relevance.

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Section: Sample Preparationmentioning

confidence: 99%

α1-Antitrypsin in serum determined by capillary isoelectric focusing

et al. 2000

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“…Because alpha-l-antitrypsin is an acute phase reactant, the serum level can vary widely depending on such factors as the presence of inflammation or the influence of estrogens, and this can result in PiMZ individuals being mistaken for normal (PiM) individuals (10). The recent use of starch-gel electrophoresis followed by crossed immunoelectrophoresis (11) and electrofocusing (12) have allowed for exact phenotyping, and have thus circumvented the problem of inadequate identification.…”

Section: Introductionmentioning

confidence: 99%

Assessment of alpha-1-antitrypsin deficiency heterozygosity as a risk factor in the etiology of emphysema. Physiological comparison of adult normal and heterozygous protease inhibitor phenotype subjects from a random population.

McDonagh¹,

Nathan²,

Knudson³

et al. 1979

J. Clin. Invest.

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A B S T R A C T For plethysmographic studies of lung mechanics and measurement of pulmonary diffusing capacity, 62 subjects were drawn from a randomly selected population sample. Data obtained from the 24 subjects of heterozygous phenotype for alpha-l-antitrypsin deficiency (PiMZ) were compared by age group with data from 38 normal (PiM) subjects matched for sex, age, and smoking history. Comparison of mean values by age group for lung volumes, diffusing capacity, lung elastic recoil, maximum expiratory flow, and the occurrence of frequency dependence of dynamic compliance revealed no differences between phenotype groups. There was no evidence of an accelerated effect of aging among PiMZ subjects when compared with normal counterparts nor was there evidence of an increased effect of smoking. From these data it appears that the PiMZ phenotype per se is not a risk factor in the development of emphysema.

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“…Für Serienuntersuchungen erscheint ein solcher Aufwand jedoch zu groß.Pierce(13) verwendete Ampholyte in dem sehr engen pH-Bereich von 4,0 bis 4,5 and 4,5 bis 5,0 und behandelte die Seren mit Reduktionsadditiva und erhielt damit eine große Auflösung. Auch Jeppsson(14) hat mit dieser Methode MjM^Subtypen untersucht.…”

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Schnelle Phänotypisierung des α 1 -Antitrypsins zur Bestimmung der Subtypen des Phänotyps MM

Bencze¹,

Sabatke²

1980

Clinical Chemistry and Laboratory Medicine

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Zusammenfassung: Die Polyaciylamid-Gel-Platten zur isoelektrischen Fokussierung wurden durch Zugabe einer bestimmten Menge Harnstoff und durch einen Vorlauf ohne Proben für eine Serummenge von 20 eingestellt. Dadurch kann eine gute Auftrennung der einzelnen Proteinfraktionen erreicht und insbesondere der Abstand zwischen der M2-und der M8-Bande bei einer Gesamtlaufstrecke von 120 mm auf über 20 mm erhöht werden. Zur Differenzierung der PiMM-Subtypen wird außerdem ein MjMj-Standardserum eingeführt und die M8-Bande in die Auswertung einbezogen. Anhand einer bereits an anderer Stelle Chest 76, im Druck) veröffentlichten klinischen Studie wird die klinische Relevanz der Methode demonstriert. Fast oi^antitrypsin phenotyping for the differentiation of PiMM-sub-typesSummary: The polyacrylamide gel slabs for isoelectric focussing were adapted for 20 samples by adding a certain quantity of urea, and by letting it run idle before adding the sample. A better separation of protein fractions was obtained, and the distance of the M2-and M8-bands was increased to more than 20 mm in a total migration length of 120 mm. A MjM 3 standard serum was introduced and the M8 band was included to differentiate PiMM sub-types. The clinical use of the method is demonstrated with the results of a recently published clinical study by Bencze, K. et al. ((1979), Chest 7(5, in press). Einführung

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α-1 Antitrypsin phenotypes determined by isoelectric focusing of the cysteine-antitrypsin mixed disulfide in serum

Cited by 67 publications

References 17 publications

α1-Antitrypsin in serum determined by capillary isoelectric focusing

α1-Antitrypsin in serum determined by capillary isoelectric focusing

Assessment of alpha-1-antitrypsin deficiency heterozygosity as a risk factor in the etiology of emphysema. Physiological comparison of adult normal and heterozygous protease inhibitor phenotype subjects from a random population.

Schnelle Phänotypisierung des α 1 -Antitrypsins zur Bestimmung der Subtypen des Phänotyps MM

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