149 results about "Bioconjugation" patented technology

The invention relates to the field of bioconjugation of functional entities, such as payloads and targeting agents, mediated by novel trans-cyclooctene (TCO) compounds. More particularly the present invention relates to novel trans-cyclooctene compounds, unnatural amino acids comprising such trans-cyclooctene compounds and preparation thereof. Said novel trans-cyclooctene compounds and unnatural amino acids comprising such trans-cyclooctene compounds are characterized by high structural stability and hydrophilicity and thus instrumental to ease the conjugation of said functional entities. The invention further relates to respective trans-cyclooctene-functionalized construct obtainable by, in particular covalently, binding said novel trans-cyclooctene compounds to a targeting agent or payload molecule, said functionalized construct containing at least one functionalizing residue comprising a mono-unsaturated trans-cyclooctene entity. The invention further relates to conjugates obtainable by covalently linking, in particular, via biorthogonal bioconjugation via a Diels-Alder-type cycloaddition, a first functionalized molecule, particularly selected from said trans-cyclooctene-functionalized constructs with a second functionalized molecule comprising a docking group (DG) capable of reacting with the trans-cyclooctene-type functional group of the first functionalized molecule. The present invention further relates to preparation methods of said constructs and conjugates. The invention also relates to the use of such conjugates of the present invention for use in medicine, to corresponding pharmaceutical compositions as well as to corresponding diagnostic and analytical kits.

Provided herein is a bio-conjugation process for preparing a composition of antibody-drug conjugates (ADCs) with improved homogeneity, wherein the antibody-drug conjugates (ADCs) with drug to antibody molar ratio of 2 (D2) are present in a high level in the composition. In the composition of ADCs prepared by the process of the present disclosure, the content of D2 is higher than 64 mol%, and the content of D0+D4+D6+D8 is less than 36 mol%, on the basis of total molar of D0, D2, D4, D6 and D8.

The invention provides a method for controlling batch-to-batch difference and improving specificity by changing raw material adding time after microsphere activation, and belongs to the technical field of biomedicine. The bioactive substances are added for coupling after the microspheres are activated for a certain time. According to the method, extra reagents or steps do not need to be introduced, batch-to-batch difference reduction and false positive reduction can be achieved only by optimizing the coupling object adding time, operation is easy, control is easy, reduction of false positive results caused by non-specific binding is facilitated, biological coupling reaction is more sensitive and reliable, and the method is suitable for industrial production. Therefore, the detection sensitivity, specificity and accuracy can be improved, and the kit has extremely high practical value in the fields which need large-scale production and have high requirements on product quality. The method provided by the invention can provide a more reliable and stable tool for biomedical research and clinical diagnosis, and has a good application prospect.

Provided herein are methods of sequencing comprising click chemistry bioconjugation. In some embodiments, target polynucleotide sequences on the same or different molecules are contacted with and hybridize to probes comprising click functional groups. Probes (e.g., reading probes) hybridizing to adaptor sequences adjacent to different sequences of interest (e.g., barcodes to be sequenced) can be hybridized simultaneously in large pools. In some embodiments, the provided methods achieve multiplexing without requiring separate hybridization of probes (e.g., reading probes) for each sequencing-by-ligation cycle, thereby reducing total hybridization time which is typically a most time-consuming step in in situ technologies. In some aspects, the hybridized probes (e.g., reading probes) are clicked onto detectable probes to analyze a sequence of a target polynucleotide in a sequencing-by-ligation fashion.

The present disclosure sets forth novel compounds and compositions including heteroaromatic silicon-fluoride-acceptors, which are useful for PET scanning. The present disclosure further includes novel methods of 18F imaging for PET scanning, the methods comprising the preparation of conjugates and bioconjugates of biological ligands of interest with heteroaromatic silicon-fluoride-acceptors. In certain embodiments the invention is practiced in the form of a kit.

Bioconjugates are described comprising alpha-fetoprotein (AFP) linked with a cytotoxin through a disulfide, glutathione-sensitive linker, for cancer treatment. In a preferred embodiment, the bioconjugate is an AFP variant linked with a maytansinoid cytotoxin via the glutathione-sensitive linker so that the bioconjugate comprises either specific high or low ratio of cytotoxins per AFP.

Methods of producing bioconjugates of O-antigen polysaccharides covalently linked to a carrier protein using recombinant host cells are provided. The recombinant host cells used in the methods described herein encode a particular oligosaccharyl transferase enzyme depending on the O-antigen polysaccharide bioconjugate to be produced. The oligosaccharyl transferase enzymes can be PglB oligosaccharyl transferase or variants thereof. Also provided are compositions containing the bioconjugates, and methods of using the bioconjugates and compositions described herein to vaccinate a subject against extra-intestinal pathogenic E. coli. (ExPEC).

A compound, or a stereoisomer, isotopomer, tautomer, or salt thereof having a structure of: A-L-B wherein A is a ligand that can covalently bioconjugate to an unnatural amino acid bearing a tetrazine or other reactive handle, incorporated into a protein; L is a linker covalently attaching A and B; and B is a ligand that can bind to a E3 ligase.

The present invention concerns a compound having the following formula (I) wherein: - m represents an integer comprised from 1 to 3, - L1 represents a bioconjugation unit, - L2 and L3 represent a spacer group, - L4 represents a release system, - L5 represents an anticancer agent, and - L6 represents a PEG moiety.

To provide a bioconjugate comprising an oligo- or polysaccharide covalently linked to a fusion protein.SOLUTION: The present invention provides a bioconjugate comprising an oligo- or polysaccharide covalently linked to a fusion protein, wherein the fusion protein comprises a ComP protein (ComP) glycosylation tag, wherein the ComP glycosylation tag comprises a cysteine residue conserved in a specific amino acid sequence, and the fusion protein is glycosylated with an oligo- or polysaccharide at a serine residue conserved in a specific amino acid sequence.SELECTED DRAWING: None

A virus conjugate comprising a genetically-modified adeno-associated virus (AAV), wherein the virus is mutated to incorporate an engineered amino acid in a site-specific manner and a genetically-modified protein of interest, wherein the protein is mutated to incorporate an engineered amino acid in a site-specific manner, wherein the engineered amino acids can further comprise orthogonal bioconjugation groups suitable for attachment of a bifunctional linking reagent, whereby the virus and protein are cross-linked via the bifunctional chemical linker reagent to form a virus conjugate is described.

Disclosed are polypeptides that can be used for bioconjugation. The polypeptides have transglutaminase activity, and they can be used to make biomolecule-compound conjugates, for example to conjugate a compound to a specific lysine or glutamine of an antibody.

Disclosed herein, inter alia, are silicon containing detectable compounds and methods of use thereof. In an aspect is provided a monovalent nucleotide or monovalent nucleoside covalently bound to a monovalent form of a compound described herein (e.g., wherein the R13 moiety of a compound described herein has reacted with a bioconjugate reactive group to form a bioconjugate linker thereby covalently bonding the monovalent compound to the monovalent nucleotide or monovalent nucleoside).

A bio-orthogonal cycloaddition reaction is provided, as well as uses in the assembly of TAC-type molecules and the preparation of bioconjugates. A method of conducting a coupling reaction includes providing a first structure comprising a diketone group or a derivative thereof and a second structure capable of providing anionic 2-furanol or a derivative thereof.

The present invention provides a bioconjugation process for preparing a composition of antibody-drug conjugates (ADCs) with improved homogeneity, wherein antibody-drug conjugates (ADCs) having a molar ratio of drug to antibody of 2 (D2) are present at high levels in the composition. In the ADC composition prepared by the method of the present invention, the content of D2 is greater than 64 mol% and the content of D0 + D4 + D6 + D8 is less than 36 mol% based on the total molar amount of D0, D2, D4, D6 and D8.

This invention discloses a selenylamide-based hydrogenation reaction of acetylacetamide compounds and its application in the modification of selenium-containing peptides and proteins, belonging to the fields of organic chemistry and chemical biology. The reaction involves the reaction of a selenium-containing compound with an acetylacetamide compound in a solvent with the addition of an additive. The applications of this reaction include: 1) selective modification and labeling of selenocysteine ​​in peptides and proteins; 2) cyclization reactions of selenium-containing peptides and selenoproteins; and 3) sequential modification of selenium-containing peptides in the presence of cysteine ​​by precise pH control combined with the hydrogen sulfide reaction of acetylacetamide. This reaction has advantages such as mild conditions, simple operation, fast reaction rate, high yield, good stability, and good selectivity. It provides a new and robust approach for the selective modification and labeling of selenium groups in bioconjugations, peptides, and / or proteins, and also provides a new method for the construction of cyclic peptides, offering a powerful tool for the development of selenium-containing peptide drugs.

The present invention relates to the field of bioconjugation of functional entities (payloads) to targeting agents, in particular biological targeting agents such as antibody drug conjugates (ADCs), in which one or more payload molecules are conjugated to a targeting agent, for example a monoclonal antibody. More specifically, the present invention relates to novel hydrophilic tetrazine molecules and their preparation, which allow more efficient conjugation of payload molecules to targeting agents such as monoclonal antibodies. The present invention also relates to certain tetrazine intermediates useful for the preparation of corresponding functionalized payload molecules. The present invention also relates to the respective conjugates, in particular bioconjugates, and methods for their preparation. The present invention also relates to the medical use of such conjugates of the present invention, to the corresponding pharmaceutical compositions, and to the corresponding diagnostic and analytical kits.

Provided for herein is a polynucleotide-modified bioconjugate comprising a substrate such as an antibody or bead linked to a conjugate component via a nucleic acid linker. Also provided are methods of making and using such bioconjugates. Conjugation methods for creating the bioconjugate are stable, not chemically harsh, and efficient enough that post-conjugation purification may not be required. Further this disclosure provides for reducing the logistic overheads related to product lines by eliminating the need for many unique linkers per conjugation pair.

The present invention relates to providing more efficient methods of conjugating two or more peptides. For example, the invention provides more efficient methods of producing antibody-drug conjugates. More particularly, the methods can be used to efficiently produce ADCs with at least two unique payloads. One approach involves the use of peptidyl asparaginyl ligases (PALs) for two consecutive ligation reactions at the N-terminal and C-terminal ends of a protein and the other approach involves the use of a chemical protein modification reaction and a PAL-mediated enzymatic ligation reaction, both able to introduce two different payloads to an antibody at specific sites.

The present invention relates to narrow emission dyes, compositions comprising the same, and methods of making and using the same. In particular, the present application provides bacteriochlorin derivatives having narrow band emission. In some embodiments, the bacteriocin derivative is PEGylated. In some embodiments, the bacteriocin derivatives have a high water solubility (e.g., 10 mg / mL or more). In some embodiments, the bacteriocin derivatives are PEGylated and have a high water solubility. The bacteriocin derivatives may comprise a bioconjugatable group for forming a conjugate (e.g., with an antibody or nanoparticle). The bacteriocin derivatives and conjugates thereof are useful for imaging and therapeutic applications. Also provided are methods of synthesizing the bacteriocin derivatives.

In general, disclosed herein are fluorogenic compounds having the structure of Formula (I):Fluorogenic compounds disclosed herein may be useful in methods for visualizing a sample. The method may include conjugating the biomolecule with sample comprising a fluorogenic compound disclosed herein; incubating the biomolecule with the fluorogenic compound for a sufficient time to allow for fluorogenic bioconjugation of the fluorogenic compound; subjecting the fluorogenic compound to an oxidative condition; contacting the fluorogenic compound with a dye; and imaging the fluorogenic compound, thereby determining the fluorescence intensity change of the fluorogenic compound.

The present invention relates to a preparation method of a photoelectrochemical sensor for detecting the transmembrane glycoprotein CD44 on the surface of breast cancer cells. The preparation method is divided into three steps. First, a TiO2 nanoarray is loaded on the surface of FTO conductive glass by a hydrothermal method. Then, a Ag nano-layer is uniformly sputtered on the surface of the TiO2 nanoarray by using a magnetron sputtering technique. Finally, the TiO2-Ag is immersed in a sodium sulfide solution to realize the local sulfidation of Ag to obtain a photoelectric conversion body, a TiO2-Ag-Ag2S nano-composite array. Then, the extraction of the transmembrane glycoprotein CD44 on the surface of breast cancer cells MDA-MB-231 is realized by using the host-guest recognition between hyaluronic acid and the transmembrane glycoprotein CD44 and a DNA strand displacement reaction. Finally, through bioconjugation and covalent bonding, the assembly of the optoelectronic material and the target on the conductive interface is realized. The preparation method of this sensor has stronger controllability compared with other reported methods, especially obtaining a sensing interface with good signal output stability. The scheme and process mentioned in this method have important reference in the fields of material synthesis, photoelectrochemical sensing, cell detection, etc.

The invention particularly relates to preparation of a CRISPR (clustered regularly interspaced short palindromic repeats)-immune combined small molecule detection probe as well as a related detection method and application thereof. The preparation method of the probe mainly adopts a chemical and biological coupling method, and the dsDNA-antibody probe is prepared by covalently coupling the dsDNA of which the 5'end is modified to NH2 and a monoclonal antibody by using a bifunctional cross-linking agent 4-(N-maleimide methyl) cyclohexane-1-carboxylic acid sulfonic acid group succinimide ester sodium salt (Sulfo-SMCC) and N-succinimido-S-acetylthioacetate (SATA). Secondly, according to the detection method, a dsDNA-antibody probe is taken as a bridge to combine an immune system and a CRISPR / Cas12a system, the specific recognition capability of an antibody to small molecules is utilized, signal amplification is carried out by virtue of RPA nucleic acid amplification, and fluorescence signals are generated by virtue of cis-cleavage activity and / or trans-single-chain DNA cleavage activity of CRISPR / Cas12a, so that conversion from immune reaction signals to high-intensity fluorescence signals is realized, and the detection sensitivity is high. And a CRISPR-immune combined small molecule detection system is established. The detection method has the advantages of simplicity in operation, high specificity, extremely high sensitivity, stable signal and the like.

A method of attaching an azide moiety to a biomolecule. The method comprises contacting a biomolecule in a solution with an azide and a copper, for a time wherein at least one azide moiety is covalently bonded to the biomolecule to yield an azidylated biomolecule. The copper is copper (I), and can be generated from copper (II) by a reductant. The solution further comprises a copper ligand for reducing degradation of the biomolecule. The azilylated biomolecule can be attached to a reagent comprising an alkyne via a copper-catalyzed azide-alkyne cycloaddition (“CuAAC”) reaction or a strain-promoted alkyne-azide cycloaddition (“SPAAC”) reaction.

The present invention provides for a metal chelator capable of binding a radioisotope or radionuclide, and bioconjugate thereof, and methods of making and using thereof.

Described herein is a general strategy to label recombinant proteins using 5HTP-directed electrochemical conjugation reaction and biomolecular conjugates produced by the covalent electrochemical reactions.

A therapeutic composition is described including a selective soluble TNF-neutralizing bioconjugate for treating TNF-mediated inflammatory disorders. The bioconjugate includes a dominant-negative TNF (DN-TNF) mutein, which contains one or more amino acid substitutions in the TNF receptor interaction domain, trimer interface domain, or both, to reduce receptor binding while enhancing heterotrimer formation with wild-type TNF, thereby neutralizing soluble TNF. The DN-TNF mutein is covalently conjugated to a biocompatible stealth polymer via a hydrolyzed maleimide linker, preventing retro-Michael reaction and polymer dissociation. The composition is formulated as an injectable buffered aqueous solution for subcutaneous, intramuscular, intravitreal, or intravenous administration. Also disclosed are methods for stabilizing the bioconjugate through controlled hydrolysis of the maleimide linker and methods for treating TNF-mediated inflammatory disorders by administering the composition to a subject in need thereof.

The invention relates to an efficient method for synthesizing self-assembling artificial proteins (SAPs) utilizing a catalytic host-guest system. The process involves encapsulating hydrophobic probes with cyclodextrin, performing site-specific protein bioconjugation, and employing a second catalysis cycle for enhanced labeling. SAPs produced through this method are versatile and find applications in therapeutic delivery and diagnostics. The process is simplified, scalable, cost-effective, and environmentally sustainable, addressing the challenges of traditional SAP synthesis. By offering improved yield, functionality, and structural integrity, this invention advances the potential of SAPs in medical and industrial fields.

The present invention provides, among other aspects, functionalized chromophoric polymer dots comprising a hydrophobic core and a hydrophilic cap, and bioconjugates thereof. Also provided are improved methods for preparing functionalized chromophoric polymer dots. Methods for in vivo imaging and molecular labeling are also disclosed.

A bio-conjugation process for preparing a composition of antibody-drug conjugates (ADCs) with improved homogeneity, i.e., a composition of antibody-drug conjugates (ADCs) with high D4 (DAR4) content through simple manipulation is disclosed, wherein the process is performed at room temperature. The resultant composition of ADCs comprises D4 in a content higher than 60 wt%, preferably higher than 65 wt%, on the basis of total weight of D0, D2, D4, D6 and D8.