Millipore Corporation

Other

Arris Pharmaceutical Corporation

University of California

Derek Hudson

Charles R Johnson

Lutz B Giebel

Ronald M Cook

Alan Frankel

Robert F Sharpe

Kevin R Shoemaker

Stewart A Thompson

Ruoying Tan

Robert T Cass

Michael J Ross

Jordan Honig

Michael Szelke

Douglas J Ng

A method is provided which facilitates and enables the production of a wide range of complex conjugates composed of similar or dissimilar units linked together by amide bonds. Said method for the production of acylthio derivatives, R&prime;&mdash;CO&mdash;SA, involves reaction of a carboxylate, R&prime;&mdash;CO&mdash;O,(or carboxylic acid R&prime;&mdash;CO&mdash;OH) with an iso-thiouronium derivative (bearing SA) in the presence of base. Nucleophilic counterion forms of the iso-thiouronium salts confer significant rate enhancement. The processes are simple, generally applicable, efficient, and do not require the employment of noxious reagents. The production of complex protein-like products by the intermediary of acylthio esters generated by the process of this invention, provides a method which is compatible with mild methods of chain assembly; and is preferably applied when the second component in the ligation bears an amino terminal cysteine residue.

Method for production of acylthio derivatives

Polyolefin particles are chemically modified by oxidation to provide a large surface area and high loading. The particles result in low back pressure in column systems, and are economical to manufacture. The particles are useful as supports in a wide range of applications including general organic as well as biopolymer synthesis, library methods, purification processes and enzyme mediated processes. In a preferred embodiment, polyethylene or polypropylene particles are oxidized in a solution containing trifluoroacetic acid or trifluoromethane sulfonic acid, chromium trioxide and sulfuric acid to provide the particles with a chemically reactive irregular surface and open channels that extend below the surface and up to essentially the length of the radius of the particles resulting in increased surface area and decreased density. The particles have pendant functional groups produced by the oxidation and/or by subsequent chemical reaction. The pendant functional groups may be attached to spacer arms that are attached to the surface of the particles. Biological material such as proteins, enzymes and whole cells may be immobilized on the particles.

Oxidized polyethylene or polypropylene particulate supports

Compositions and methods for identifying compositions which inhibit binding of the Rev protein to the Rev-responsive element in cells infected by HIV-1 are provided. The compositions display or mimic the electronic configuration of a binding domain within the native Rev protein, but are free from those portions responsible for REV activity and are preferably modified to display enhanced binding affinity to the RRE. Screening methods for identifying polypeptides and other compositions having such enhanced binding affinity are also described.

Compositions and methods for inhibiting replication of human

Method and apparatus for simple and rapid preparation of reusable, addressable surface-immobilized arrays of biomolecules (libraries) used for screening for interaction with any biologically significant target. A special plate having on its surface a plurality of discreet functionalized substrate areas, typically in arrays of 10.times.10 to 400.times.400, is provided for chemical synthesis or bonding thereon of desired families of biomolecules (e.g. peptides, DNA, RNA, oligosaccharides). In the case of peptides, such as hexapeptides, the resulting permanently hexapeptide-loaded plate is a reusable Addressable Synthetic Peptide Combinatorial Library (ASPCL), in which 1 to 3 (typically two) of the positions in the sequence are uniquely identified by the address location. Plate embodiments include substrates of physically bonded (e.g. glued) conventional particulate materials such as Pepsyn-K, or functionalizable films of linear or crosslinked polymers covalently attached to, or physically adhered to the plate surface. Spacer arm moieties may also be attached to the substrates. A unique multi-slot block assembly is used to prepare the ASPCLs. In library applications, for example determining peptides which bind to functional proteins (enzymes, receptors, antibodies), the substrate-bound peptides are assembled with several positions consisting of uniformly distributed equimolar mixtures of residues, and 2 separated or sequential positions uniquely identified by their spatial location on the substrate array, the 'address'. Following identification of the known residues giving the greatest affinity for the arrayed positions in the sequence, optimal binding for the complete peptide sequence is determined by an iterative process replacing formerly mixed positions with known AAs at unique addresses.

Method and apparatus for peptide synthesis and screening

Method and apparatus for simple and rapid preparation of reusable, addressable surface-immobilized arrays of biomolecules (libraries) used for screening for interaction with any biologically significant target. A special plate having on or in its surface a plurality of discreet functionalized substrate areas, typically in arrays of 10.times.10 to 400.times.400, is provided for chemical synthesis or bonding thereon of desired families of biomolecules (e.g. peptides, DNA, RNA, oligosaccharides). In the case of peptides, such as hexapeptides, the resulting permanently hexapeptide-loaded plate is a reusable Addressable Synthetic Peptide Combinatorial Library (ASPCL), in which 1 to 3 (typically two) of the positions in the sequence are uniquely identified by the address location. The preferred plate embodiment employs an HPMP wink of porous polyolefin removably received in holes in the plate. A unique multi-slot block assembly is used to prepare the ASPCLs. The wink carrier plate is also employed with a vacuum block system to assist in washing, deprotection, and probing. In library applications, for example determining peptides which bind to functional proteins (enzymes, receptors, antibodies), the substrate-bound peptides are assembled with several positions consisting of uniformly distributed equimolar mixtures of residues, and 2 separated or sequential positions uniquely identified by their spatial location on the substrate array, the 'address'. Following identification of the known residues giving the greatest affinity for the arrayed positions in the sequence, optimal binding for the complete peptide sequence is determined by an iterative process replacing formerly mixed positions with known AAs at unique addresses.

Pilot apparatus for peptide synthesis and screening

The invention relates to methods and systems of unhindered construction and display of tethered organic ligand molecules, and more particularly to preparation and use of thin film, substantially non-crosslinked hydrophilic polar multi-functionalized polymers (HPMPs) anchored to a variety of functionalized substrates so that the HPMP forms a thin film matrix layer providing a unique highly hydrated, high dielectric environment equivalent to an aqueous solution, for affinity binding of Ligands (L) to Tagged Target Molecules (TTMs). Ligands, and especially MER.sub.n ligand libraries such as peptide libraries, are singly tethered to the HPMP by a 'permanent' strong covalent bond so that subsequent displacement of the TTM does not also displace the ligand from the HPMP, thereby making the HPMP tethered Ligand library reusable. The HPMP thin film is on the order of 200-2000 .ANG. thick, is highly accessible (to the TTMs), and permits flexible, 3-D display of the singly tethered ligands for free permeability therein of the TTMs for affinity binding. The 3-D nature of the HPMP film provides high amplification of display, and the open nature of the matrix permits rapid elution of excess TTMs and other molecules. Importantly, the HPMP matrix layer, while providing amplification is also non-masking, being essentially transparent to detection methods. The combination of the reuseability, amplification and non-masking properties results in a very significant, many-fold increase in sensitivity and speed of screening assays.

Thin film HPMP matrix systems and methods for constructing and

Derivatives of 1-phenyl pyrazolin-5-one have many applications for biomolecule synthesis. Enol esters of protected amino acids made by the present process provide efficient coupling in solid-phase peptide synthesis. The 1-phenyl-pyrazolin-5-one derivatives are highly crystalline, stable, non-toxic and easy to prepare. Many possess self-indicating properties, facilitating spectrophotometric monitoring and automation of peptide synthesis.

Active esters for solid phase peptide synthesis

A chemical processing system is disclosed for the automated dissolution, dispersing and reaction of chemicals, especially for synthesizing proteins. The system includes a plurality of storage cartridges containing a first chemical in fluid communication with a reservoir containing a second chemical to be reacted with the first chemical. Each cartridge includes a pump (e.g., a plunger) which operates by changing the internal volume of the cartridge. The pump permits bi-directional flow of the first chemical into and out of the reservoir and cartridge to promote mixing and reacting with the second chemical in the reservoir to produce a third chemical.

Apparatus and process for performing repetitive chemical processing

Trialkoxybenzyl (Taob) protected asparagine and glutamine, a method of synthesis and a method of use are provided. The Taob protected Asn and Gln have the following formulae: ##STR1## wherein Z is an alkyl group having from 1 to 10 carbon atoms; X and W are any .alpha.-protecting group which can be selectively removed while maintaining Taob intact; Y is H or any group sufficiently active or activatable to react with NH.sub.2 -- or NH.dbd. to generate an amide bond; n is 1 for asparagine or 2 for glutamine. These derivatives are stable in solution, have good solubility in organic solvents and couple directly without side reactions.

Protecting groups for asparagine and glutamine in peptide synthesis

A novel method of activation of a protected amino acid using Benzotriazolyloxytris(dimethylamino)phosphonium Hexafluorophosphate (BOP) as an activator for solid phase peptide synthesis is provided. Hydroxybenzotriazole is included in preferred embodiments. An intermediate acyloxyphosphonium salt is formed from the protected amino acid and the BOP which immediately reacts with HOBt to form an ester without formation of the less reactive symmetrical anhydride. A tertiary or quaternary substituted nitrogen compound must be used to produce a carboxylate of the protected amino acids as it also functions to increase their solubility.

BOP reagent for solid phase peptide synthesis

Compounds corresponding in structure to enkephalin or polypeptide analogues thereof, wherein one or more peptide links of the enkephalin or analogue is represented by a group or groups the same or different selected from dimethylene, methylene-imino and keto-methylene groups.

Growing community of inventors

San Anselmo, CA, United States of America

Derek Hudson