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Biomedical subjects

K Sagisaka

Publications and source records attributed to K Sagisaka.

At least 91 records · Page 5Linked to original sources
PubMed1981

Human specific antigens and species difference in non-hemoglobin proteins of red cell lysate.

Non-hemoglobin proteins (NHPs) of 12 species of mammalian were effectively isolated from red cell lysates with CM-Sephadex chromatography. Polyacrylamide gel disc electrophoresis demonstrated that human and monkey NHPs had 13 and 11 bands, respectively, but guinea pig and swine two to three bands. Rabbit anti-human NHP reacted with monkey, bovine, canine and badger as well as human. Absorption of the anti-NHP with monkey NHP resulted in preparing human specific anti-NHP. The specific antiserum gave two bands locating near the origin in immunoelectrophoresis. Human specific antigens were prepared by affinity chromatography with the anti-NHP. The antigens were composed of three bands, two of which had slightly faster mobility than carbonic anhydrase and the other migrated fastly on disc electrophoresis. These bands were proved to have neither LDH nor esterase activity.

Animals↗
PubMed1980

Application of precipitating activity of Canavalia lineata DC lectin for isolation of alpha 2-macroglobulin from human plasma.

To isolate alpha 2-macroglobulin from human plasma, complex of Canavalia lineata DC lectin and plasma was used as a starting material. The complex was solubilized with 0.08 M phosphate buffer pH 7.2 containing 0.9% NaCl and 0.02 M alpha-methyl-D-mannoside. The solution was applied onto a Bio gel A-5m column and eluted with solubilizing buffer. Of three peaks, only the second peak contained alpha 2-macroglobulin. After equilibrating with 0.06 M potassium phosphate buffer pH 6.8, the fraction was applied onto a hydroxylapatite column and eluted with the same buffer. The eluate was confirmed to be extremely pure alpha 2-macroglobulin; immunoelectrophoresis with anti-human serum produced a single precipitation line, and SDS disc electrophoresis gave a single band at alpha-globulin region. Few differences in carbohydrate composition between this preparation and those prepared by other methods were observed. By this method, about 45% of alpha 2-macroglobulin present in plasma was recovered.

Blood Protein Electrophoresis↗
PubMed1980

Individual difference in non-hemoglobin proteins of red cell lysate.

Non-hemoglobin proteins (NHP) were effectively isolated from red cell lysate by CM-Sephadex chromatography using 0.01 M phosphate buffer pH 6.3. NHP from 180 subjects was analyzed with polyacrylamide gel disc electrophoresis. The individual difference was observed at the intermediately migrating zone after protein staining. Group I had two of the three bands migrating quickly, group II all the three bands, and group III two of the three bands migrating slowly. Incidences of groups I, II and III were 67, 20 and 13 percent, respectively.

Adult↗
PubMed1980

An improved isolation procedure for carbonic anhydrase C from red cell lysate.

An effective isolation of carbonic anhydrase C (CA-C) from red cell lysate was described. The lysate dialyzed against 0.0175 M phosphate buffer pH 6.3 was applied onto a CM-Sephadex column equilibrated with the same buffer. The elution was performed with the starting buffer and 0.1 M dibasic potassium phosphate containing 0.14 M NaCl. The hemoglobin fraction eluted with the second eluant was applied onto a DEAE-cellulose column and eluted with 0.2 M glycine containing 0.01 percent KCN, resulting in complete isolation of CA-C at high recovery rate. For the preparation in a large scale, the hemoglobin fraction prepared from CM-Sephadex semi-batch-type chromatography was treated with cold ethanol and chloroform. The purity of these preparations was confirmed by polyacrylamide gel disc electrophoresis and immunoelectrophoresis.

Animals↗
PubMed1979

Preparation of anti-alpha 2-macroglobulin using Canavalia lineata DC lectin for differentiating species-specificity of blood stains.

To differentiate species-specificity of blood stains, anti-alpha 2-macroglobulin was raised in rabbits against Canavalia lineata DC lectin-serum complex (LSC). Adsorption of anti-LSC with human lipoprotein resulted in antiserum specific for alpha 2-macroglobulin. It was confirmed by Ouchterlony test that the antiserum adsorbed successively with monkey serum or anti-LSC adsorbed directly with monkey serum reacted with only human serum but not with mammalian ones. Immunoelectrosyneresis and anti-LSC consumption test could identify species-specificity of blood stains kept for up to two years and for up to several years, respectively. It is indicated that anti-LSC is quite effective for differentiating species-specificity of blood stains.

Adsorption↗
PubMed1979

On the receptors of human red cells reacting with Phaseolus coccineus L. lectin.

In order to elucidate receptors of proteolytic enzyme-treated red cells which react with Phaseolus coccineus L. lectin, the receptors prepared by affinity chromatography were serologically investigated. P. coccineus lectin had high agglutinin activity for bromelin-, papain- and pronase-treated red cells but that for the cells treated with ficin and trypsin was relatively low. Analyses of chemical composition revealed that sialic acid of the receptors from normal red cells was considerably much as compared with that from the treated cells. On the contrary, the enzyme treatment did not affect particularly carbohydrate composition of the receptors. Disc electrophoresis showed that the patterns of receptors from red cells treated with bromelin or papain were different from those from the other cells. On two-dimensional immunoelectrophoresis, the receptor of trypsin-treated cells gave five precipitation lines against anti-stroma and that of papain-treated cells three lines, but any other receptors showed no line. These findings indicate that there are plural receptors for P. coccineus lectin in red cells treated with each of proteolytic enzymes and that the receptors from respective red cells have electrophoretically and serologically different property.

Chromatography, Affinity↗
PubMed1978

Fragmentation of human albumin with proteolytic enzymes and its antigenicity with special reference to human-specificity.

Human albumin prepared by the trichloroacetic acid method was treated with bromelin, ficin, papain, pronase or trypsin. Pronase- or trypsin-treated albumin showed four fragments on polyacrylamide disc electrophoresis. When rabbit antiserum to pronase-treated albumin was adsorbed with monkey albumin, antibody with human specificity was completely abolished, whereas anti-albumin serum adsorbed with pronase-treated albumin retained the antibody to human albumin. It was considered that antigenic site with human specificity in albumin structure was inactivated by pronase.

Antigens↗
PubMed1977

Preparation of anti-M and -N antibodies by use of dextran gel as an immunoadsorbent.

Anti-M and -N antibodies are usually prepared from rabbit immunized with human O.M or O.N red cells. The crude antisera must be adsorbed with non-corresponding red cells to remove heteroagglutinin. To specify the crude antisera, we have employed dextran gel (DEAE-Sephadex A-50) as an immunoadsorbent, which was mixed with M or N group substance prepared by the phenol-saline method, and was fixed with formaldehyde. The substance-coupled gel was added to the corresponding crude antiserum and washed with cold saline. Then, the gel added with an equal volume of saline was heated at 55 degrees C for 10 min and applied on a funnel with slight evacuating. By this method, specific anti-M and -N antibodies (titer 1:16 to 1:32) were prepared, but the antibody was not eluted from the gel reacted with IgM fraction. The recovery rate and specificity of the eluted antibodies are discussed.

Dextrans↗
PubMed1977

Comparative studies of non-hemoglobin proteins of adult and newborn red blood cell lysates.

It has been known that hemolysate is composed of not only hemoglobin but also non-hemoglobin proteins (NHP). NHP isolated effectively from hemolysate of adult and newborn by CM-Sephadex chromatography was investigated by polyacrylamide disc or slab electrophoresis. It was demonstrated on slab electrophoresis that adult NHP is composed of 4 zones (approximately 14 bands) with different mobilities and that adult NHP shows a considerably different pattern from cord NHP. The difference was observed also on the electrophoresis stained for esterase or lactate dehydrogenase. Rabbit antiserum to adult NHP was adsorbed with cord NHP and specific antiserum for adult NHP was prepared. However, antiserum reacting with only cord NHP was not obtained from anti-cord NHP serum. These results suggested that adult specific components with passable antigenicity were present in adult NHP and that cord NHP had similar antigenicity as adult NHP.

Adult↗
PubMed1976

On the agglutinogens of red cells developed with proteolytic enzymes and neuraminidase.

It has been known that the agglutinability of human red cells is changed or enhanced by treatments with proteolytic enzymes or neuraminidase. In this paper, the serological properties of agglutinogens developed by proteolytic enzymes (bromelin, ficin, papain, trypsin and pronase) and neuraminidase are investigated by using antisera to trypsin- and neuraminidase-treated red cells. The adsorptions of the antiserum to trypsinized red cells with the cells treated with each of the proteolytic enzymes showed that the agglutinogens uncovered by bromelin, ficin and papain were different from those by pronase and trypsin. It was demonstrated that pronase was the most effective enzyme to uncover the agglutinogen located on deeper site of red cell membrane. This was confirmed by the agglutination with the test cells treated twice with two kinds of the enzymes. The reactions of the antiserum to neuraminidase-treated red cells treated with six kinds of the enzymes indicated that the agglutinogens developed by neuraminidase resembled those by bromelin, ficin and papain more than those by trypsin and pronase.

Agglutinins↗
PubMed1976

Incidence and quantitation of pregnancy-associated globulin in the Japanese.

It has been pointed out that a serum protein specific for pregnancy (pregnancy-associated globulin, PAG) appears in females near term and is maintained during some period after delivery. The incidence and quantitation of PAG in 120 pregnant females and normal subjects, 1,469 males and 355 females, were studied by double diffusion and single radial immunodiffusion methods. All the pregnant females were proved to have PAG. The incidence of PAG in normal males and in females over 16 years of age was 5.3% and 36.9%, respectively. In the females, the incidence of PAG tended to increase with aging; 63% over 31 years of age and 100% over 51 years. Quantitative analyses, which were expressed as a percentage of the pooled pregnant sera, showed that PAG OF pregnant sera ranged from 24 to 240 and that of normal sera from 1 to 33.

Adolescent↗
PubMed1976

A simple method to remove hemoglobin from antiserum.

Hemoglobin contamination in antiserum occurs frequently in immunohematology. The hemoglobin obscures the ring test for the observation of antigen-antibody reaction. A simple method to remove hemoglobin from antiserum is described in this paper. An appropriate amount of CM-Sephadex was added to antiserum containing hemoglobin and the mixture was packed into a cellulose tube, followed by dialysing against 0.01 M monobasic sodium phosphate solution (pH approx. 6). Equilibrating with stirring, the hemoglobin was completely adsorbed with CM-Sephadex so that hemoglobin-free antiserum was prepared without dilution of the antiserum. This method is useful for immunohematology.

Hematology↗
PubMed1975

One step separation of the sialoprotein possessing ABH and MN blood group activities from human red cell membrane.

Sialoprotein possessing blood group activity has been usually extracted with organic solvents from red cell membrane. To obtain the blood group substance under milder condition, the author presented carboxymethyl (CM) cellulose column chromatography with eluents of phosphate buffer containing 1% Brij 35. Red cell stroma was applied onto the column with an equal volume of 0.0175 M phosphate buffer pH 6.3 and one-fifth volume of 0.05% 2-mercaptoethanol in 1% Brij 35. Separation of the substance was performed by stepwise elution with three kinds of phosphate buffers. Serological assay showed that only the first fraction eluted with 1% Brij 35 in 0.0175 M phosphate buffer pH 6.3 and ABH and MN blood group activities and the other fractions obtained with buffers of higher molarities had not such activities. Sialic acid was also detected only in the first fraction, and the majority of proteins of the stroma was found in the other fractions.

ABO Blood-Group System↗