Recombinant Fragment of the Extracellular Domain of Human Desmoglein 3 Fused with the Fc-Fragment of Human IgG1 Selectively Adsorbs Autoreactive Antibodies from the Sera of Pemphigus Patients.

Using the recombinant second fragment of the extracellular domain (EC2) of human desmoglein type 3 (Dsg3) as an affinity ligand, an immunosorbent was obtained that selectively binds autoreactive antibodies to this domain from the immune sera of patients with pemphigus. The EC2 protein was obtained in the form of a fusion protein with the Fc-fragment of human IgG1. The production was carried out in CHO cells using the method of transient expression.

Application of an eremomycin-chiral stationary phase for the separation of DL-methionine using simulated moving bed technology.

Recently a new chiral stationary phase (CSP) was introduced, based on the immobilization of the macrocyclic glycopeptide eremomycin to epoxy-activated silica. The application of this new CSP to preparative enantioseparation using simulated moving bed (SMB) chromatography will be presented. MeOH-H(2)O (0.1M NaH(2)PO(4))=20/80 (v/v) was used as the mobile phase to separate the enantiomers of methionine. Successful separation was realized providing productivities around 15 g(product)/l(stat)/h for both l and d-methionine under nonlinear conditions. In such delicate continuous chromatographic separation processes, besides productivity, the long-term stability of the applied stationary phases is of importance. Column to column fluctuations were negligible and long-term stability of the preparative stationary phase was satisfactory according to the results of perturbation experiments performed before and after long-term SMB runs.

[Sorbents with immobilized glycopeptide antibiotics for separating optical isomers by high-performance liquid chromatography].

Chiral sorbents for HPLC separation of optical isomers carrying glycopeptide antibiotics (eremomycin or its eremosaminyl aglycon, ristomycin, or vancomycin) fixed onto the surface of silica gel have been synthesized. The patterns of the retention and separation of profen isomers and their dependence on the nature of the chiral selector and the eluant composition have been studied. The sorbents were shown to be highly enantiospecific in separating the isomers of alpha-amino-, alpha-hydroxy-, and alpha-methylphenylcarboxylic acids (profens).

New chiral stationary phase with macrocyclic glycopeptide antibiotic eremomycin chemically bonded to silica.

A new chiral stationary phase (CSP) was prepared by attachment of macrocyclic glycopeptide antibiotic eremomycin to the epoxy-activated silica under mild conditions. In contrast to CSP with immobilized vancomycin, which is a close structural analogue of eremomycin, the prepared CSP reveals high enantioselectivity for separation of amino acids enantiomers. It was demonstrated by the example of ristocetin A CSP that method of the immobilization of macrocyclic glycopeptide antibiotics affects remarkably the resulting enantioselectivity.

[Chromatographic properties of human leukocyte alpha-interferon A on sorbents with sepharose and silochrome matrices]. / Khromatograficheskie svoistva chelovecheskogo leikotsitarnogo alpha-interferona A na sorbentakh s sefaroznymi i silokhromnymi svoistvami.

Chromatography on immobilized monoclonal antibodies NK-2 from a bacterial strain-producer resulted in a pure human leukocyte alpha-interferon A (alpha-INF-A) homogeneous upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and reverse phase high performance liquid chromatography. The chromatographic properties of partially purified alpha-INF-A on synthetic and commercial sorbents containing immobilized dyes, aromatic dipeptides, chelating and hydrophobic ligands as well as on porous glass have been investigated. In most cases, [125I]alpha-INF-A was used as an inner standard. The chromatographic behaviour of native and [125I]-labeled alpha-INF-A was practically the same. alpha-INF-A was most effectively chromatographed on porous glass, L-Trp-L-Trp-Sepharose 4B and Cu2+-chelate sorbents. In the latter case, the feasibility of substitution of the Sepharose matrix for the silochrome one has been demonstrated. It has been proposed that alpha-INF-A has a hydrophobic "pocket" with exposed aromatic amino acid residues which are capable of selective binding to aromatic dipeptides.