Tandem purification of mouse IgM monoclonal antibodies produced in vitro using anion-exchange and gel fast protein liquid chromatography

A tandem chromatographic procedure was used to isolate rapidly mouse IgM monoclonal antibodies produced by cultivation of hybridomas in vitro. Hybridoma culture supernatants containing mouse IgM monoclonal antibodies were first chromatographed on an anion-exchange Mono Q column connected to a fast protein liquid chromatography system. This anion-exchange step offers the advantage of obtaining IgM antibodies in a concentrated form. The IgM-rich fractions from the Mono Q column were then injected on a gel filtration Superose 6 column equilibrated with a low-ionic strength buffer and eluted with a high-ionic strength buffer. Assessment of the purity of isolated IgM monoclonal antibodies was performed by sodium dodecyl sulphate polyacrylamide gel electrophoresis together with a Coomassie Brillant Blue R 250 staining technique. Assessment of the immunoreactivity of isolated IgM monoclonal antibodies was performed by an enzyme linked immunosorbent assay using a solid phase adsorbed antigen against which IgM monoclonal antibodies were directed. The chromatographic procedures described allows the rapid isolation of mouse IgM monoclonal antibodies produced in vitro at a high degree of purity and in an immunoreactive state.     

Ring-chain tautomerism of substituted hydrazones

The freshly distilled products of the reaction of acetaldehyde with 2-(N-propylhydrazino)- and 2-(N-isopropylhydrazino)ethanols have substituted perhydro-1,3,4-oxadiazine structures and are isomerized to the corresponding N-alkyl-N-(2-hydroxyethyl)hydrazones via an equilibrium. Mixtures of both tautomeric forms were isolated by the reaction of these hydrazino alcohols with acetone and pinacolone.See [1] for communication II.Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 9, pp. 1167–1170, September, 1971.     

Azines and azoles. 64. Mass spectra and tautomerism of 2-aryl-4,6-dioxo-1,3-thiazines

Mass spectra of 2-aryl-4,6-dioxo-1,3-thiazines were studied and the general fragmentation paths of these compounds determined. Based on mass spectra of compounds with fixed structures, 2-aryl-5,5-dialkyl-4,6-dioxo-1,3-thiazines, 4-alkoxy- and 4-acyloxy-2-aryl-6-oxo-1,3-thiazines, the decomposition paths of the dicarbonyl and hydroxycarbonyl forms were determined. It was concluded on the basis of the obtained data that 2-aryl-4,6-dioxo-1,3-thiazines exist in the gas phase predominantly in the hydroxycarbonyl form.For Communication 63, see [1].Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 10, pp. 1409–1412, October, 1987.     

Über einige Fraktionierungen laugenlöslicher Anteile von Kunstfaser-Zellstoffen

Zusammenfassung Aus Fichten- und Buchen-Kunstfaserzellstoffen wurden -und -Cellulosen präparativ hergestellt und diese polymeranalog nitriert. Die entstandenen Nitrate wurden durch Behandeln mit Aceton in einen löslichen und unlöslichen Teil getrennt, die gewichtsmäßig bestimmt wurden. Die acetonlöslichen Nitratanteile wurden aus den Acetonlösungen durch steigende Wasserzusätze fraktioniert gefällt und die Fraktionen gewichtsmäßig bestimmt; weiters wurden die reduzierten Viskositäten der Fraktionen bestimmt. Während die Stickstoffgehalte der acetonlöslichen Nitratanteile etwa jenen von Cellulosenitraten entsprechen, betragen die Stickstoffwerte der acetonunlöslichen nur etwa die Hälfte bis ein Drittel davon. In den acetonlöslichen Anteilen liegen im wesentlichen Hexosane vor. Die höchsten Fraktionen der -Cellulosen enthalten Anteile bis annähernd etwa DP 200. Im Gegensatz dazu bestehen die acetonunlöslichen Nitratanteile von - und -Cellulosen hauptsächlich aus Mischungen von Nitraten von Pentosanen und Polyuronsäuren. Die Acetonlöslichkeit von Nitraten laugenlöslicher Anteile aus Kunstfaserzellstoffen gibt demnach orientierende Hinweise auf die chemische Zusammensetzung dieser Anteile.Herrn Prof. Dr.A. Wacek zum 60. Geburtstag gewidmet.Die vorliegenden Untersuchungen wurden bereits in den Jahren 1952 und 1953 durchgeführt.     

1,2,4-Triazine and triazole derivatives from α-keto acids and thiosemicarbazides

Thiosemicarbazides of 2-hydroxyphenylglyoxylic acid, which, depending on the conditions, undergo cyclization to interconvertible 5-(2-hydroxyphenyl)-6-oxo-1,2,4-triazine-3-thiones or (2-hydroxy-benzoyl)-²-1,2,4-triazoline-5-thiones, are formed by the action of thiosemicarbazide and its homologs on coumarindione. The thiosemicarbazones of 2-hydroxyphenylglyoxlyic and other 2-substituted phenylglyoxylic acids undergo cyclization to 6-(2-hydroxyphenyl)-5-oxo-1,2,4-triazine-3-thiones.Deceased.Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 1, pp, 116–121, January, 1977.     

Formylation of 2-phenyl- and 3-phenyl-5-hydroxybenzofuran derivatives

The Vilsmeier formylation of 2-phenyl- and 3-phenyl-5-hydroxybenzofuran derivatives was studied. It is shown that 2-phenyl-5-methoxybenzofuran is formylated in the 4 position, whereas 3-phenyl-5-methoxybenzofuran is formylated in the 2 position.Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 8, pp. 1023–1024, August, 1976.     

Determination of metal traces in tantalum by proton activation analysis

Procedures for determining traces of calcium, titanium, vanadium, chromium, iron, copper, niobium, molybdenum and tungsten in tantalum are described utilizing 11 MeV proton activation. The instrumental method of analysis with the use of a Ge(Li) detector and the radiochemical separation of the corresponding fractions are described. The formula for the calculation of the concentration of traces with the use of thick target yields is given. Experimental data on the thick target yields of radioisotopes⁴⁴Sc,⁵²Mn,⁵⁶Co,⁶⁵Zn,^93mMo,⁹⁶Tc are presented. Based on the experimental data, the detection limits are estimated at 1–10 ppb for the above mentioned metal traces in tantalum. The successful solution of the task of the simultaneous determination of nine elements in tantalum confirms the wide applicability of proton activation analysis.     

Schiff bases in syntheses of substituted naphthylamines,naphthyridines, azaphenanthrenes,and benzocarbazole

9-Methylnaphtho[2,1-b][1,6]naphthyrldine and 2,3-dimethyl-1-azaphenanthrene were obtained on a K-16 dehydrogenating catalyst from N-(1,2,5-trimethyl-4-piperidylidene)--naphthylamine, while N-cyclohexylidene--naphthylamine yielded N-phenyl--naphthylamine and benzo[a]carbazole.Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 10, pp. 1400–1403, October, 1972.     

The chemistry of indole

The opening of the pyrazolidine ring at the nitrogen-nitrogen bond in 1-arylpyrazolidine under the influence of acidic agents which leads to the formation of N-(3-aminoalkyl)indoles is described. The method enables one to obtain indoles with branched aminoalkyl radicals.For part XIII see [1].     

The alkylation of benzimidazole derivatives with γ-valerolactone

Reaction of 1, 3-dimethylbenzimidazolone and its 5-methyl and 5-chloro derivatives with -valerolactone in presence of anhydrous aluminum chloride gives the corresponding 6-(1-methyl-3-carboxypropyl)-benzimidazolones.