Ensayo inmunoenzimático para la determinación de anticuerpos naturales anti banda 3 / Immunoenzimatic assay to determine the natural anti-band 3 antibodies

Se estandarizó un método inmunoenzimático (ELISA) para la determinación de anticuerpos naturales anti banda 3 acoplando a placas Chemobond la proteína banda 3 obtenida a partir de la membrana de eritrocitos humanos mediante una combinación de cromatografía de intercambio aniónico y afinidad. Se emplearon 3 mg de la proteína banda 3 por pozo, lográndose un nivel de detección de 1 mg/mL de anticuerpo anti banda 3. Los resultados indican la utilidad del método para su empleo en la medición de los niveles de anticuerpos naturales anti banda 3 en pacientes con drepanocitosis, tanto en estado basal como en las crisis vasooclusivas dolorosas.

An immunoenzimatic assay (ELISA) was standardized to determine the natural anti-band 3 antibodies by coupling with Chemobond plates the band 3 protein obtained from the membrane of human erythrocytes by a combination of affinity and anion exchange chromatography. 3 g of band 3 protein per well were used. A level of detection of 1 g/mL of anti-band 3 antibody was achieved. The results showed the usefulness of the method to measure the levels of natural anti-band 3 antibodies in patients with drepanocytes, both in basal state and in the painful vasocclusive crises.

Erythrocyte membrane proteins in healthy Saudis and patients with hereditary spherocytosis and hereditary elliptocytosis

Little is known about hereditary spherocytosis [HS] and hereditary elliptocytosis [HE] in the native population of Saudi Arabia, even though these conditions are seemingly common. The purpose of this study was to ascertain the protein make-up of the red cell membrane in healthy Saudis and in patients with HS and HE. Patients and Eighteen healthy Saudi subjects [13 males and 5 females], 11 patients with HS [6 males and 5 females] and 11 patients with HE [7 males and 4 females] were studied. All normal controls and patients underwent SDS-PAGE red cell membrane protein analysis in duplicate and the stained protein bands were identified and quantitated by densitometry. In normal, healthy Saudis, the mean values for seven membrane proteins [alpha spectrin, spectrin, ankyrin, band 3, protein 4.1, protein 4.2, and actin] were similar to those published for normal, healthy Americans. Of the eleven cases with HS, 7 [64%] demonstrated detectable protein abnormalities while 4 [36%] were apparently normal. The electrophoretic patterns of membrane proteins in Saudis with HS differed from those of patients with HS in other parts of the world. Of the 11 cases of HE, 7 [64%] displayed abnormalities while 4 [36%] were normal. The electrophoretic pattern of the main proteins in the membranes of red blood cells in healthy Saudis is similar to that reported from the USA. However, significant differences exist in the electrophoretic patterns between Saudi patients with HS and patients from other parts of the world

Abnormalities of erythrocyte membrane proteins in Korean patients with hereditary spherocytosis

Hereditary spherocytosis (HS) is a common inherited erythrocyte membrane disorder characterized by chronic hemolytic anemia. Clinical manifestations and biochemical abnormalities of HS are heterogeneous. In this study, we investigated erythrocyte membrane protein defects in 27 Korean HS cases. Utilizing both the Fairbanks system and the Laemmli system, sodium dodecyl sulfate polyacrylamide gel electrophoresis of erythrocyte membrane proteins was performed. Proteins were stained with Coomassie brilliant blue and gels were scanned using a densitometer. We detected spectrin deficiency in 7.4% of cases (2/27), ankyrin deficiency in 29.6% (8/27), combined spectrin and ankyrin deficiency in 3.7% (1/27), band 3 deficiency in 11.1% (3/27) and protein 4.2 deficiency in 14.8% (4/27). Membrane protein deficiencies were not observed in nine cases (33.3%, 9/27). Members of two of seven families tested showed the same protein defects as the proband. Ankyrin deficiency alone and combined with spectrin deficiency accounted for 33.3% of cases (9/27), and they were the most common biochemical defects in Korean HS cases. Protein 4.2 deficiency caused HS more frequently in Koreans than in Caucasians.

Abnormalities of erythrocyte membrane proteins in Korean patients with hereditary spherocytosis

Hereditary spherocytosis (HS) is a common inherited erythrocyte membrane disorder characterized by chronic hemolytic anemia. Clinical manifestations and biochemical abnormalities of HS are heterogeneous. In this study, we investigated erythrocyte membrane protein defects in 27 Korean HS cases. Utilizing both the Fairbanks system and the Laemmli system, sodium dodecyl sulfate polyacrylamide gel electrophoresis of erythrocyte membrane proteins was performed. Proteins were stained with Coomassie brilliant blue and gels were scanned using a densitometer. We detected spectrin deficiency in 7.4% of cases (2/27), ankyrin deficiency in 29.6% (8/27), combined spectrin and ankyrin deficiency in 3.7% (1/27), band 3 deficiency in 11.1% (3/27) and protein 4.2 deficiency in 14.8% (4/27). Membrane protein deficiencies were not observed in nine cases (33.3%, 9/27). Members of two of seven families tested showed the same protein defects as the proband. Ankyrin deficiency alone and combined with spectrin deficiency accounted for 33.3% of cases (9/27), and they were the most common biochemical defects in Korean HS cases. Protein 4.2 deficiency caused HS more frequently in Koreans than in Caucasians.