Clontech Laboratories, Inc.

Paul S Nelson

Te-Tuan Yang

Thomas H Smith

Steven R Kain

Mark A Kent

Brian Perry

Purification of poly-amino acid-tagged recombinant proteins has been improved by the use of a carboxymethylated aspartate ligand complexed with a third-block transition metal having an oxidation state of 2and a coordination number of 6. A method for synthesizing the metal ion-CM-Asp complex is also described. Further, the metal ion-CM-Asp complex can be used for screening protein function.

Method for purification of recombinant proteins

Purification of poly-amino acid-tagged recombinant proteins has been improved by the use of a carboxymethylated aspartate ligand complexed with a third-block transition metal having an oxidation state of 2,and a coordination number of 6. A method for synthesizing the metal ion-CM-Asp complex is also described. Further, the metal ion-CM-Asp complex can be used for screening protein function.

Purification of poly-amino acid-tagged recombinant proteins has been improved by the use of a carboxymethylated aspartate ligand complexed with a third-block transition metal having an oxidation state of 2.sup.+ and a coordination number of 6. A method for synthesizing the metal ion-CM-Asp complex is also described. Further, the metal ion-CM-Asp complex can be used for screening protein function.

The subject invention concerns novel reagents and methods useful for introducing functional groups and labels at multiple sites of a nucleotide. The reagents are useful, for example, in constructing labeled oligonucleotide hybridization probes for detecting the presence of a target polynucleotide in biological or chemical samples.

Non-nucleoside 1,3-diol reagents and their use to label or modify

A trifunctional reagent possessing a primary hydroxyl, a secondary hydroxyl, and a primary amino group. This reagent is useful in solid phase oligonucleotide synthesis for the convenient labeling of the 3'-terminus. The secondary hydroxyl may be an phosphoramidite derivative permitting the attachment to the solid phase support. The reporter molecule may be attached to the trifunctional molecule prior to the completion of the oligonucleotide synthesis or after the oligonucleotide is cleaved from the support. A preferred species of this trifunctional molecule is shown below.

Non-nucleoside 1,3-diol reagents for labeling synthetic oligonucleotides

A method for derivatizing and labeling the 3'-terminus of an oligonucleotide during solid phase synthesis comprising the use of a multifunctional reagent whose preferred structure is shown below. ##STR1## wherein CPG is controlled pore glass beads, Fmoc is 9-fluorenylmethyoxycarbonyl, and the alkylamine contains 1 to 50 carbon atoms.

Method for labeling the 3' terminus of a synthetic oligonucleotide using

The subject invention concerns a novel multifunctional solid support reagent which is useful in solid phase oligonucleotide synthesis. Specifically, the reagent is used in a solid phase oligonucleotide process to chemically modify the 3' terminus of a synthetic oligonucleotide with any chemical functional group. The invention reagent can be used to construct 3' labeled oligonucleotide hybridization probes to detect the presence of a target polynucleotide in biological and clinical samples.

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Union City, CA, United States of America

Paul S Nelson