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Summary and conclusions1. Serum from 6 patients who had received radioiodide or I131-labeled thyroxine was studied by zone electrophoresis in filter paper, at pH 8.6. Most of the I131 had a mobility intermediate between the alpha1- and alph2-glabulins, and a small proportion had the mobility of albumin. No differences were detected between thyroxine added to serum (in vivo) and the thyroid hormone secreted by a normal thyroid gland, by a Graves' disease thyroid gland, and by a functioning thyroid carcinoma. 2. Two patients had received very large doses of radio-iodide and their serum contained large amounts of butanol-insoluble I131, whereas in the other patients the serum I131 was largely butanol soluble, and behaved like thyroxine on paper chromatography. These two forms of serum I131 had similar mobility on zone electrophoresis, but could be differentiated by fractional precipitation of the serum proteins with phosphate buffer. The radioiodine, therefore, was probably attached, in each instance, to a d...

1.1. Filter paper electrophoretic analysis of the serum and urine proteins in thirty-five cases of multiple myeloma has been carried out. In all thirty-five cases characteristic protein abnormalities were observed; seventeen cases showing homogeneous peaks in both the serum and urine, eleven cases in serum only, seven cases in urine only. 2.2. All urine samples showing a positive Bence-Jones protein reaction also displayed a characteristic electrophoretic abnormality. Eight of the twenty-four electrophoretically positive urine samples were Bence-Jones protein negative by the usual tests. Because of the relative simplicity of urine electrophoresis by the filter paper technic, this test is considered to be a practical clinical procedure. 3.3. Two cases are presented in detail because certain atypical clinicopathologic features of their disease patterns possibly assist in an understanding of the relationship of the plasma cell dyscrasias to chronic lymphatic leukemia and the lymphosarcomas. A third case, which demonstrates the value of serial electrophoretic studies in myeloma, is also presented. 4.4. Consideration is given to some of the physicochemical properties of the myeloma serum and urine proteins, and to the possible nature of their interrelationship.

A simplified procedure for filter paper electrophoresis is described in which disturbing factors such as evaporation, heating, buffer concentration gradients, and pH changes in the electrode vessels were reduced to a minimum. Artificial mixtures of highly purified proteins could be separated and the components isolated. The application of the method to a variety of studies on serum proteins is demonstrated. Protein concentration in paper segments was determined by two different methods of protein estimation. Curves were obtained showing the same five major peaks for normal serum as found by the classical methods of free electrophoresis. Comparisons were made of the areas of the various components under the curves obtained with the different methods. Two dimensional electrophoresis was applied to serum and serum components. It proved of value in demonstrating the heterogeneity of fractions such as the γ-globulin of serum. The polysaccharide dextran was used as an index of the extent of electro-osmotic flow during the course of the various experiments. The ratio of the distance of electroosmotic flow and the distance of protein migration was shown to be constant for a given type of paper. For serum albumin on Munktell 20 paper this ratio was 0.35. A formula for mobilities applicable to liquid in a highly porous supporting medium is presented. Mobility values for human serum albumin at various pH levels on paper showed approximate agreement with those obtained in free solution giving a similar isoelectric point.

SummaryNine serum protein fractionation experiments using the continuous flow paper electrophoresis technique were performed. Six experiments were performed using the serum collected from individual patients 24 and 48 hours after receiving therapeutic doses of I-131 and 3 using the serum of euthyroid individuals which was incubated in vitro for 1/2 hour at 37°C with I–131 labelled thyroxine prior to fractionation. Protein fractions obtained were identified by conventional paper electrophoresis and analyzed for radioactivity, total protein and cholesterol, the latter being taken as an index of lipid contents. Similar results were obtained for both in vivo and in vitro experiments. Beta globulins were found to be capable of forming thyroid hormone protein complexes to a larger extent than gamma globulins, but the largest amount of radioactive hormone was found as an alpha-2 globulin-thyroid hormone complex. A patient showing a marked abnormality in his plasma proteins showed the highest percent of radioacti...

Filter paper electrophoretic analyses of serum were carried out in a series of eighteen proved cases of multiple myeloma, twenty-three normal controls and 130 patients with non-myelomatous diseases, and compared with the results of classical Tiselius electrophoresis and chemical fractionation. Abnormal electrophoretic patterns were found in all of the cases of multiple myeloma studied, and tall narrow spikes representing abnormal proteins with varying mobilities in the globulin range were demonstrated in 78 per cent. The latter were considered diagnostic of myelomatosis since similar patterns have not been described in any other disease and since none were observed in any of the present controls. In six of the eighteen cases the diagnosis was first established on the basis of these electrophoretic patterns and subsequently confirmed by bone marrow biopsy. In eight others they were merely confirmatory in previously diagnosed cases. Of special interest were the diagnostic patterns found in four patients in whom myelomatosis was not suspected. There were significant quantitative differences between the results obtained by filter paper and classical electrophoresis, and the results of both differed from those of chemical fractionation. However, the diagnostic peaks of myelomatosis were equally well demonstrated by both electrophoretic methods. It is concluded (1) that the electrophoretic patterns of serum obtained by both the filter paper and Tiselius methods exhibit highly characteristic peaks in a high proportion of individuals with multiple myeloma, (2) that such patterns are of great diagnostic value and (3) that the filter paper method offers the advantages of simplicity, low cost and speed, without any loss in sensitivity or specificity, which make it especially valuable for screening large numbers of specimens in patients suspected of having myelomatosis.

CHANG (1955) reported the lowering of the blood hemoglobin of chicks due to free gossypol feeding and postulated that free gossypol in the diet leads to interference with the synthesis of the proteins of the blood in these chicks. In view of the useful information that has been obtained in recent years using the electrophoresis technique in the study of normal and pathological sera, an experiment was planned to study the electrophoretic patterns of serum samples obtained from birds fed a normal diet, as well as from birds fed free gossypol in the diet. An electrophoresis cell using filter paper strips, as originally designed by Durrum (1950) and marketed by Spinco, was employed. Later it was discovered that the resolution of the globulins of chick serum is very poor on filter paper. More serum samples were obtained from birds used in a study of the effect of protein level on …

In few conditions is the electrophorogram of serum proteins capable alone of deciphering the diagnosis. In conjunction with the assembled clinical data and relevant applied pathological tests the definition of the disease is often clarified. The patterns as observed by the applied pathologist are serially discussed and the importance of the variations in different fractions presented. In several instances, exemplified by liver disease, the assistance rendered toward prognosis has been considerable. Perhaps the most clearcut diagnosis is possible in the agammaglobulinaemia syndrome which is immediately evident by the technique of paper electrophoresis. Current advances are mainly in the direction of estimating the protein-bound carbohydrate and lipid constituents which are believed to reflect the abnormalities in several infections and metabolic diseases. Utilisation of non-ionic detergents has also allowed separation of subfractions of the globulins into numerous new components, including lipo-proteins, some of which had been previously observed during the application of improved buffer systems designed to restrict interference of heavy metals and to facilitate accentuation of fractions having intermediate mobility.

A reproducible technic is presented for the staining of lipoproteins on paper. This method gives a greater total uptake of dye than previously described procedures, especially by the α-lipoproteins. The staining is accomplished with a solution of Sudan Black B in a mixture of methyl and isopropyl alcohols and the excess dye is removed by wash solution of 30% isopropanol.

(1952). Reproducibility with Paper Electrophoresis of Serum Proteins. Scandinavian Journal of Clinical and Laboratory Investigation: Vol. 4, No. 4, pp. 307-312.

An application of the biuret reaction to the determination of protein fractions on filter paper electropherograms of serum is described. The relative mobilities of the serum protein fractions of the domestic fowl and of man are compared. Values are reported for serum protein fractions as separated by filter paper electrophoresis in a methanolic veronal buffer. Some observations on the serum proteins of the estrogenized immature pullet are reported; and it is tentatively suggested that another fraction as well as serum phosphoprotein appears in the serum of the pullet as a consequence of treatment with estrogen.

SummaryThe use of blood plasma as a buffer for filter paper electrophoresis makes it possible to study the physico-chemical state of injected materials or of native blood constituents without subjecting the blood fractions to possible alterations by an artificial buffer. In veronal buffer yttrium appeared to exist in wholly immobile forms. In plasma buffer, yttrium existed in an immobile fraction and in an unionized fraction which moved with the solvent. Migration of materials as a result of solvent flow must be considered as a factor in the interpretation of electrophoretic studies of this type.

Investigation of various steps involved in separation of serum proteins by paper electrophoresis and their quantitation by dyeing with bromphenol blue has revealed that the results obtained are quite dependent on a number of variables. It is essential, therefore, that rigid control of these variables be exercised and that normal values be obtained by the specific technic adopted. Normal values have been given for the technic employed and data on stability of sera at room and refrigerator temperatures have been presented.

An investigation of the electrophoretic data from patients with various malignant diseases shows that the possibilities of paper electrophoresis as a diagnostic tool are limited. The relative inaccuracy of the method may cover up important details; but more physiological information can scarcely be expected from data concerning only one physical property of a protein molecule. Quantitative evaluation of the data still gives rather qualitative results. With more material it may be possible to differentiate between various groups of diseases. The use of the “entropy” of a serum protein mixture does not give greater differentiation. Only in a limited number of cases can it be used to distinguish pathological and normal sera. The high correlation in the normal group between the albumin contribution to the total “entropy” and the total “entropy” provides a criterion for the detection of pathological serum protein mixtures in most cases.

I investigated amino acids composition of the protein from the tuberculous pus plasma in 3 cases and free amino acids in the tungstic acid filtrates of tuberculous pus plasma by a paper-chromatographic procedure. 1) I found tuberculous pus plasma proteins do not essentially differ from usual proteins regarding the qualitative composition. 2) In 3 cases of closed tuberculous abscess never punctured free amino acids were not detected. 3) Free amino acids were present in the protein-free filtrates of tuber-culous pura of 5 closed abscesses often punctured before. 4) The number of the free amino acids found there, however, far less than that of the corresponding amino acids in hot true pura. There is a striking contrast between tubercuous and non-tuberculous true pura.

Summary The electrophoretic pattern of serum proteins has been studied in 30 infants suffering from toxicosis. During the stage of acute dehydration the main changes were a moderate or marked increase of alpha globulin, particularly of the α2 fraction and a decrease of albumin. In some cases there was also an increase of the gamma globulin. During recovery the alpha globulin decreases while albumin increases. However, the return to normal of the electrophoretic pattern is not always achieved when clinical recovery occurs. The absolute amounts of total serum proteins have also been estimated and their significance has been discussed. Electrophorese sur papier des proteines du serum au cours de la toxicose. L'aspect electrophoretique des proteines du serum a eteetudie chez 30 ehfants presentant une toxicose. Au cours de la periode de deshydratation aigue, on a pu noter une augmentation nette des alphaglobulines (en particulier la fraction alpha 2), et une diminution de l'albumine. Dans quelques cas, on a pu noter aussi une augmentation des gammaglobulines. La diminution des alphaglobulines, et l'augmentation de l'albumine est survenue avec la guerison. Neanmoins, le retour a la normale de la courbe d'electrophorese n'est pas toujours acheve au moment de la guerison elinique. Papierelektrophorese von Serum-Protein bei Sauglingstoxikose. Die Elektrophoresebefunde von Serumproteinen sind bei 30 an Toxikose leidenden Sauglingen untersucht worden. Wahrend des Stadiums der akuten Dehydration waren die Hauptveranderungen eine massige oder ausgepragte Zunahme des Alpha-Globulins, insbesondere der α2-Fraktion und eine Abnahme des Albumins. In einigen Fallen be-stand auch eine Zunahme des Gamma-Globulins. Wahrend der Genesung nimmt das Alpha-Globulin ab, wahrend das Albumin ansteigt. Jedoch ist der Ruckgang zum nor-malen elektrophoretischen Bild nicht immer vollendet wenn die klinische Genesung eintritt. Electroforesis en papel de las proteinas sericas en la toxicosis. Se estudiaron las imagenes electroforeticas de las proteinas sericas en 30 lactantes afectos de toxicosis. Durante el periodo de deshidratacion aguda, los cambios mas notables fueron un moderado o marcado aumento de la alfa globulina, especialmente de alfa 2. Durante la convalecencia, la fraecion alfa globulina disminuyo mientras au-mentaba la albumina. Sin embargo, la vuelta a la imagen electroforetica normal no siempre es completa al ocurrir, la mejoria clinica.

The radioisotopes of yttrium, cerium, promethium and ytterbium were administered intravenously as chlorides to rats. The blood serum was collected after 5 and 60 min respectively. Furthermore these radioisotopes, as chlorides, were added to rat serum in vitro. The binding of the rare earths to the serum proteins was studied by means of paper electrophoresis, free columnar electrophoresis in a sucrose gradient at various pH values, and gel filtration through Sephadex at various pH values. It was found that these elements had already combined with albumins and low-molecular u.v. absorbing complexes in the serum 5 min after injection. No differences were observed between the results obtained when the isotopes were added to serum in vitro or injected intravenously.

Plasma and sera from normal and leukemic (erythroblastosis) chickens have been studied with the aid of paper electrophoresis. Differences are more easily demonstrable with pH 6.4 phosphate buffer than with barbiturate buffer. There is a relative depression in serum albumin and an apparent 'shift' in the positions normally occupied by the α- and β-globulins. The latter regularly appear to move as one band in electropherograms of sera from leukemic birds. It has been possible to induce this 'shift' in normal sera by storage or incubation.

A comparison of serum protein fractions (electrophoretic separation) between fed female control and alloxan diabetic rats indicates a decrease in percentage albumin and an increase in percentage α2-, ß- and γ-globulins. Total protein also decreased and so did the grams percentage albumin and α1-globulin, while the grams percentage ß-globulin rose. In the depancreatized series there was a decrease in both percentage and grams percentage albumin and α1-globulin and an increase in the percentage and grams percentage ß- and γ-globulins. With the exception of a rise in γ-globulin, sham operated rats showed no change in any category. Control male rats have higher α1 and lower γ-globulin values than female rats, but both male and female rats show the same changes following the production of diabetes by either alloxan administration or partial pancreatectomy. Fasting control or diabetic rats 24 hours produces no change in the serum protein patterns despite the disappearance of observable lipemia during the fast.

Carboxymethylcellulose (CMC) was used to control electroosmosis during paper electrophoresis. Increasing the negative charge on the matrix with CMC reduced the adsorption of certain lipoproteins and negatively charged substances. Advantage of these effects was taken to study the appearance of α-lipoproteins in calf serum and to separate them from other components either with or without the use of surface-active agents. The further use of CMC in curtain separations is indicated.

Summary In 21 premature infants, the birth weights of whom ranged between 1500 and 2400 g and the ages between 1 and 56 days, the total serum protein values have been determined and the serum protein components have been electrophoretieally differentiated. All infants were fed on human milk but, in 10 infants additional protein was administered (Casein preparation Secoosan). Total protein values were found to be lower in premature infants than in full-term infants. During the first eight weeks of life total protein values decreased, from 5.63 g per cent to 4.03 g per cent. The decrease in total protein is caused by a decrease in the amount of albumin (from a little below 3 g per cent in the first week of life to a little above 2 g per cent in the eighth week) and of gamma-globulin (from about 1 g per cent to about 0.5 g per cent) while the alpha1-, alpha2-and beta-globulin values remain rather constant. If casein is added to the diet the total serum protein values and the serum albumin values will range at higher levels, but the decrease with increasing age will not be prevented. On the contrary, additional casein to the diet will cause a further reduction in the gamma-globulin concentrations (Fig. 1). Taux des proteines dans le serum des prematures. Des determinations du taux des proteines totales ainsi que des taux particuliers des differentes proteines ont ete effectuees par electrophorese chez 21 bebes nes avant terme. Tous ces enfants etaient nourris au lait maternel et 10 d'entre eux recurent des supplements de proteines. Il est apparu que le taux des proteines totales etait plus faible chez les prematures que chez les enfants nes a terme. Au cours des huit premieres semaines qui suivirent la naissance, le taux des proteines totales descendit de 5,63 g%a 4,03 g%. Cet abaissenient du taux des proteines totales provenait de la diminution de la concentration en alburnine (qui d'abord legerement inferieure a 3 g % au cours de la premiere semaine etait descendue a un niveau legerement superieur a 2 g % au bout de huit semaines) et en gamma-globuline (qui, etant d'environ 1 g %etait descendue a environ 0,5 g %), tandis que les taux des globulines alpha1, alpha2 et beta etaient restes relativement constants. L'addition de caseine a la nourriture fit monter les taux des proteines totales et de l'albumine a des niveaux plus eleves, mais ces concentrations diminuerent egalement avec le temps. D'autre part, l'addition de caseine a la nourriture entraina une diminution plus importante du taux des gamma-globulines. Serumproteinwerte bei fruhgeborenen Kmdern. Bei 21 fruhgeborenen Kindern wurden die gesamten Serumproteinwerte bestimmt und die Serumproteinbestandteile elektrophoretisch differenziert. Alle Kinder wurden nut Frauenmilch ernahrt, aber 10 Kinder erhielten einen Eiweisszusatz. Die gesamten Proteinwerte erwiesen sich bei den fruhgeborenen Kindern als niedriger als in vollreif geborenen. Wahrend der ersten 8 Lebenswochen fielen die totalen Proteinwerte von 5,63 % auf 4,03 % ab. Der Fall im Gesamtproteingehalt ist durch einen Abfall im Albumingehalt bedingt (von ein wenig unter 3 g % in der ersten Lebenswoche auf ein wenig uber 2 g % in der achten Woche) und Gammaglobulingehalt (von ca. 1 g % auf ca. 0,5 g%), wahrend die Alpha1-, Alpha2-und Betaglobulinwerte ziemlich konstant bleiben. Wenn Kasein zur Diat zugefugt wird, werd. en die gesamten Serumproteinund Serumalbuminwerte ein hoheres Niveau erreichen, aber der mit dem Alter einhergehende Abfall wird nicht verhindert werden. Im Gegenteil, der Kaseinzusatz zur Diat bewirkt eine weitere Herabsetzung der Gammaglobulinkonzentrationen. Valores seroproteicos en los prematuros. Han sido determinados, en 21 prematures, los valores seroproteicos totales y diferenciados electroforeticamente los componentes seroproteicos. Todos los ninos fueron alimentados con leche humana, pero a 10 de ellos se les administraron proteinas adicionales. Hallose que los valores proteicos totales eran inferiores en los prematuros a los de los ninos nacidos a termino. Durante las ocho primeras semanas de la vida, los valores proteicos totales disminuyeron. de 5.63 g % a 4.03 g %. La disminucion del total proteico esta causada por una disminueion de la cantidad de albumina (desde un poco menos de 3 g % en la primera semana de la vida hasta un poco mas de 2 g % en la octava semana) y de gammaglobulina (desde aproximadamente 1 g % a aproximadarnente 0.5 g %) mientras los valores globulinicos alfa1, alfa2 y beta, permanecen bastante constantes. Si se anade caseina a la dieta, los valores seroproteicos totales y los valores seroalbuminosos alcanzaran niveles mas elevados, pero la disminueion, al aumentar la edad, no podra evitarse. La caseina, contraria, adioionada a la dieta, causara todavia mas reduccion en las concentraciones gammaglobulinicas.

Seromucoids from sulphosalicylic acid filtrates of normal and pathological sera were concentrated by means of tannin and caffeine. The concentrated seromucoid was separated by paper electrophoresis into 5 fractions with mobilities corresponding to those of the total serum. The level of seromucoid in normal and pathological sera and the levels of the seromucoid electrophoretic fractions were determined by the tannin micromethod. Limits of normal ranges of individual seromucoid fractions and their content in serum proteins were established. High variability of the level of the electrophoretic seromucoid fractions was found in pathological sera.

A method is described for use of a volatile triethylamine-acetic acid buffer for continuous-flow paper electrophoresis of human plasma. Separation is excellent and contamination is minimal. The properties of the buffer are well suited for additional analytical studies.

A method is proposed for the measurement of corticosteroid-binding globulin in human serum. The method is easy to carry out and necessitates only 2 ml of serum. The number of binding sites present in the plasma proteins with an electrophoretic mobility between the α1 and α2 globulins is selectively measured.

312 male chronic alcoholists without clinical symptons of cirrhosis hepatis are inoculated with heat-killed typhus-paratyphus bacilli. The result of the inoculations is compared with the serum proteins of the patients. A connection between the O-antigens and the γ-globulins is established, and also between specific and unspecific H-antigens and the (α-) β-globulins.

1. 1. Tiselius's nomenclature of electrophoretic fractions ( α 1 , α 2 , α 3 , β 1 , β 2 , γ ) can be applied to the description of serum protein fractions of the grass-snake. 2. 2. β-Globulins prevail on electrophoretic strips, in proteinograms and in the quantitative distribution of serum proteins of the grass-snake. 3. 3. α-Globulins are the second largest component of the protein spectrum after β-globulins. 4. 4. The seasonal variation in the serum protein pattern reveals itself in a “regrouping” in the quantitative distribution of all electrophoretic fractions.

濾紙電気泳動法を家畜の臨床面に応用するための基礎的資料を得る目的から, 諸種の臨床検査の結果, 健康と判定した多数例の牛・馬・豚・緬羊・山羊・犬・猫および家兎について, 同一の泳動条件下に血清蛋白の濾紙電気泳動分析を実施した. その成績は, いずれも平均値をもって示した. この研究の前半においては, 健康家畜の血清蛋白の"正常値"に関して, 文献的考察を試みるとともに, 年令による変動を検討した. その結果, 幼若動物と成熟動物との間に差があることを認め, また同一種の中でも, 品種によって若干の差異がみられることを明らかにした. さらに後半では, 犬の"正常値"の概念を明確にするため, 発育および年令の増加にともなう血清蛋白の動態を精細に検討するとともに, 品種・性・飼養条件・日差・運動ならびに妊娠・分娩の影響について検索した. その結果,末梢血中の microfilaria が陰性で, 臨床上健康と認められる雑種犬多数の平均値を, 一応, 健康犬の"正常値"と見なして差支えないことを知った. 稿を終わるに当たり, 終始懇篤な御指導と御校閲を賜わった大越伸教授に衷心より謝意を表する. また, 実験遂行上, 絶大な援助をいただいた内科教室員各位に深謝する. さらに, 材料の採取に種々便宜を与えられた関係各位に厚く御礼申し上げる.

SummaryRats which have developed parabiosis intoxication show, in addition to the other hematologic aberrations which characterize the condition, changes in the blood proteins. Anemic partners have a low plasma protein level and polycythemic animals an elevated plasma protein titre. The partitions which albumin and the respective globulins contribute toward the total serum proteins average the same in both anemic and polycythemic twins, although as compared to control levels there is a diminished serum albumin and an elevated α2-globulin component in each.

The development of rapid and dependable methods of paper electrophoresis for fractionation of serum proteins has brought a valuable contribution (1) to diagnostic laboratory procedures. Electrophoretic analysis in the pemphigus group of diseases has been made in the search of yet They generally agreed that in the acute form and in the extensive and severe cases of the disease the total protein and the albumin were decreased and the gamma globulin was increased. The results for the other fractions showed some discrepancies. The chronic or mild cases were TABLE I Number Age (years) Duration of Disease Last Shower of Bullae Sedimentation Rate (Wester- gren-mm/ hour) General Condition Albumin (%) Globulins Total Protein Albu-of Patients Alpha-l (%) Alpha-2 (%) Beta (%) Gamma (%) G?ob ulin 1 23 4 ys.