39 results about "Glycan biosynthesis" patented technology

The current invention provides polypeptides and polypeptide conjugates that include an exogenous N-linked glycosylation sequence. The N-linked glycosylation sequence is preferably a substrate for an oligosaccharyltransferase (e.g., bacterial PgIB), which can catalyze the transfer of a glycosyl moiety from a lipid-bound glycosyl donor molecule (e.g., a lipid-pyrophosphate-linked glycosyl moiety) to an asparagine (N) residue of the glycosylation sequence. In one example, the asparagine residue is part of an exogenous N-linked glycosylation sequence of the invention. The invention further provides methods of making the polypeptide conjugates that include contacting a polypeptide having an N-linked glycosylation sequence of the invention and a lipid-pyrophosphate-linked glycosyl moiety (or phospholipid-linked glycosyl moiety) in the presence of an oligosaccharyltransferase under conditions sufficient for the enzyme to transfer the glycosyl moiety to an asparagine residue of the N-linked glycosylation sequence. Exemplary glycosyl moieties that can be conjugated to the glycosylation sequence include GlcNAc, GlcNH, bacillosamine, 6-hydroybacillosamine, GalNAc, GaINH, GlcNAc-GlcNAc, GlcNAc-GlcNH, GlcNAc-Gal, GlcNAc-GlcNAc-Gal-Sia, GlcNAc-Gal-Sia, GlcNAc-GlcNAc-Man, and GlcNAc-GlcNAc-Man(Man)2. The transferred glycosyl moiety is optionally modified with a modifying group, such as a polymer (e.g., PEG). In one example, the modified glycosyl moiety is a GIcNAc or a sialic acid moiety.

Compositions and methods for producing compositions comprising immunoglobulins or immunoglobulin fragments having an N-linked glycosylation pattern consisting predominantly of the GlCNAcMan₃GlcNAc₂ N-glycan structure are disclosed. The GlCNAcMan₃GlcNAc₂ N-glycan structure effects an increase in binding to the FcγRiπ receptors and a decrease in binding to the FcγRH receptors.

Methods and materials are provided for the production of compositions of erythropoietin protein, wherein said compositions comprise a pre-selected N-linked glycosylation pattern as the predominant N-glycoform.

The present invention relates to the recombinant method used for the production of a highly glycosylated form (in total five N linked glycosylations as opposed to three N linked glyosylations in the natural EPO) of erythropoietin. The added sites for glycosylation will result in greater number of carbohydrate chains, and higher sialic acid content than human EPO, which in turn would impart to the recombinant molecule a longer half-life. The invention further relates to the construction of expression cassettes comprising nucleic acid sequences encoding for the highly glycosylated form of Erythropoietin and stable expression in the host cells. The invention further relates to the optimized method for purification of the erythropoiesis stimulating protein. The recombinant EPO according to the invention, and the salts and functional derivatives thereof, may comprise the active ingredient of pharmaceutical compositions for an increase in the hematocrit for treatment of anemia and for restoration of patient well being and quality of life.

The invention relates to glycoengineered antibodies and antibody-conjugates. In particular, the invention relates to an antibody conjugate, prepared from an IgG antibody comprising one N-linked glycosylation site on the combination of a single heavy chain and single light chain, wherein the N-linked glycosylation site is a mutant N-linked glycosylation site as compared to its wild type counterpart. The invention further relates to methods for the preparation of the antibody-conjugates according to the invention.

E2 is one of the three envelope glycoproteins of Classical Swine Fever Virus (CSFV). E2 is involved in several functions including virus attachment and entry to target cells, production of antibodies, induction of protective immune response in swine, and virulence. Seven putative glycosylation sites in E2 were modified by site directed mutagenesis of a CSFV Brescia infectious clone (BICv). A panel of virus mutants was obtained and used to investigate whether the removal of putative glycosylation sites in the E2 glycoprotein would affect viral virulence/pathogenesis in swine. We observed that rescue of viable virus was completely impaired by removal of all putative glycosylation sites in E2, but restored when mutation N185A reverted to wild-type asparagine produced viable virus that was attenuated in swine. Single mutations of each of the E2 glycosylation sites showed that amino acid N116 (N1v virus) was responsible for BICv attenuation. N1v efficiently protected swine from challenge with virulent BICv at 3 and 28 days post-infection suggesting that glycosylation of E2 could be modified for development of CSF live-attenuated vaccines. Additionally, a new developed virus, contained deletions of putative glycosylation sites N1 in E2 and N1 in E0 (6b), called N1E0/2v, induce a solid protection against the challenge at 3 and 28 days post-inoculation.

The invention discloses a trans-N-glycosidase BtNGT, which is derived from bibersteinia trehalosi, and the amino acid sequence is shown as SEQ ID NO.1. The invention further discloses an application of the trans-N-glycosidase BtNGT in carrying out N-linked glycosylation modification on polypeptide, the trans-N-glycosidase BtNGT can serve as a tool enzyme for polypeptide glycosylation modification to simply and rapidly utilize UDP-Glc or UDP-Gal to glycosylate polypeptide containing N-X-S/T sequence, other glycosyltransferases can then be utilized to modify the carbohydrate chain of glycopeptide, and thereby a target glycopeptide is obtained. A new approach to the glycosylation modification of polypeptide and protein is provided, and a simple method is provided for the synthesis of glycoprotein vaccines.

The invention encompasses among other things antibodies including cytotoxic antibodies such as anti-CD20 having avian N-linked glycosylation patterns obtained from egg white of eggs laid by transgenic avians.

The present invention provides a pharmaceutical composition useful for treating bacterial infections in humans and animals which comprises administering to a human or animal in need thereof, an antibacterially effective combination of a β-lactam antibiotic and an inhibitor of any bacterial peptidoglycan biosynthesis enzyme, especially GlmU, GlmU, MurA, MurB, MurC, MurD, MurE, MurF, MurG, MraY, and UppS. Further provided is a method of discovering synergists for antibiotics including: a) expressing in a cell an antisense nucleic acid against a nucleic acid encoding a gene product so as to reduce the activity or amount of the gene product in the cell, thereby producing a cell sensitized to an antibiotic; b) characterizing the sensitization of the cell to the antibiotic and selecting pairs of antibiotics and genes that result in antibiotic efficacy at one-fifth or less the concentration required in the absence of the antisense gene; c) screening for chemical compounds that inhibit the gene product corresponding to the selected synergistic gene; and d) selecting or creating chemical analogs that inhibit the gene product corresponding to the selected synergistic gene such that the inhibition occurs in the bacteria.

The present invention relates to a method of diagnosing colorectal cancer by detection of glycan changes, and more particularly to a method of diagnosing colorectal cancer using mass spectrometry, in which, when specific glycan structures increase, decrease or significantly change due to a change in N-linked glycosylation of a colorectal cancer patient-derived glycoprotein, as detected by mass spectrometry, the glycan structures are selected as diagnostic markers.

There is disclosed a pharmaceutical composition for treating solid tumors that overexpress HER-2, comprising an agent selected from the group consisting of (a) an isolated polypeptide having from about 50 to 79 amino acids taken from the sequence of SEQ ID NO. 1, wherein the polypeptide binds to the extracellular domain ECD of HER-2 with an affinity binding constant of at least 10⁸ M⁻¹, (b) an isolated and glycosylated polypeptide having from about 300 to 419 amino acids taken from the sequence of SEQ ID NO. 2, wherein the C terminal 79 amino acids are present, and wherein at least three N-linked glycosylation sites are present, (c) a monoclonal antibody that binds to the ECD of HER-2, and (d) combinations thereof, with the proviso that the agent cannot be the monoclonal antibody alone, and a pharmaceutically acceptable carrier.

This invention provides a quantitative analysis of glycan biosynthesis along meta pathways using computer simulation for comparing a computer generated spectrum to experimental data to quantitatively track the biosynthesis. Computer simulating the mass spectra of isotopic detection of aminosugars with glutamine experiments allows modeling the glycan biosynthesis over time, via changes in the ¹⁴N and ¹⁵N isotope abundance levels, to estimate the relative abundance of molecules involved in glycan biosynthesis, from experimental mass spectra. Gradient search optimization is used to maximize the coefficient of determination between the experimental spectrum and the simulated spectrum. These relative abundances are then fed into a pathway simulation model to analyze glycan biosynthesis. Simulating a mass spectrum allows reconfirming the identification, quantifying the isotopic configurations and obtaining the relative abundance of each as samples are taken at periodic intervals, which is then organized to allow tracking the changes in abundance levels over time.

This invention relates to a method for gastric cancer diagnosis through the detection of glycan changes, and to a kit for gastric cancer diagnosis. More specifically, based on the fact that in gastric cancer patient-derived haptoglobin, there are changes in N-linked glycosylation of haptoglobin, which are detected through lectin and mass spectrometery, that is, an increase in fucosylation, increases or significant changes in specific glycan structures depending on the classification of antennary structures, or a remarkable decrease in a high mannose structure of the N-glycan as compared to normal persons, N-glycan structures identified using the changes in N-linked glycosylation of haptoglobin may be usefully used as a diagnosis marker in a method for gastric cancer diagnosis using lectin or mass spectrometry, and a kit for gastric cancer diagnosis.

Compositions and methods for producing aglycosylated plasminogen (PLG) polypeptides and fragments and variants thereof are provided. Compositions of the invention include isolated nucleic acid molecules encoding aglycosylated PLG polypeptides in which the asparagine (Asn) residue corresponding to residue Asn-289 of the mature human PLG polypeptide has been substituted with an amino acid residue that does not support N-linked glycosylation at that position of the PLG polypeptide, as well as the aglycosylated PLG polypeptides encoded thereby. Expression constructs comprising these PLG-encoding nucleic acid molecules and transgenic plants comprising these expression constructs are also provided. Methods of the invention comprise introducing a PLG-encoding nucleic acid molecule of the invention into a plant of interest and culturing the plant under conditions to produce the aglycosylated PLG polypeptide. The aglycosylated PLG polypeptide allows for significant increases in production and yield of PLG from a plant-based expression system without comprising the ability of the PLG product to be activated to a polypeptide capable of binding fibrin and having serine protease activity, including biologically active plasmin that is also glycosylated. The activated aglycosylated plasmin is useful to treat diseases or conditions associated with a thrombus.

Mullerian Inhibiting Substance analogues are disclosed which comprise one or more non-naturally occurring N-linked glycosylation sites, protease cleavage sites, and/or tags such as an epitope tag. Also disclosed are pharmaceutical compositions comprising such compositions and methods of using such compositions, for example in the treatment of conditions associated with uncontrolled growth of a tissue derived from a Mullerian duct, such as the condition endometriosis.

The invention relates to modified Factor VIII molecules with reduced N-linked glycosylation and reduced immunogenicity. The invention also relates to methods of using modified Factor VIII molecules, for example, to treat patients afflicted with hemophilia.

The invention discloses application of shenling baizhu powder in preparing a drug for treating TNBS-induced colitis. The shenling baizhu powder can improve the richness of an effective microbial flora, reduce the abundance of a harmful microbial flora and has a certain reduction effect on the flora diversity at the same time to achieve the flora balance of the whole intestinal environment. The shenling baizhu powder also has certain auxiliary effects on the recovery of IBD, and can also increase the abundance of various metabolic pathways and restore carbohydrate metabolism, lipid metabolism and glycan biosynthesis and metabolism of the intestinal effective microbial flora to make the metabolism normalized and alleviate the adverse effects of TNBS-induced rat IBD on the intestinal flora ofthe organism from both the flora structure and the metabolic function. The shenling baizhu powder is a traditional Chinese medicine composition which is more natural than other therapeutic drugs andhas no side effects.