Ricki M Helm

6441142, Aug 27, 2002

Jun 18, 1999

09/336463

Ricki M. Helm - Little Rock AR

University of Arkansas - Little Rock AR

C07K 1600

5303879, 53038885, 435331, 4353441

Peanuts are a common cause of food hypersensitivity reactions. The sera of 10 patients who had atopic dermatitis and a positive double-blind placebo-controlled food challenge to peanut were used to investigate the major allergens of peanut. Crude Florunner extracts were fractionated by anion-exchange chromatography using a step gradient (limit buffer, 0. 05M BisTris/1. 5M NaCl). One hundred microliters of each 2. 0 ml fraction was dot-blotted onto nitrocellulose paper and IgE-binding activity assessed using the serum pool to select allergen-containing fractions. A protein peak (OD 280) which eluted at 10% NaCl and demonstrated intense IgE-binding was further analyzed by two-dimensional SDS-PAGE/immunoblot analysis. The majority of this fraction is a protein which has a molecular weight of 17 kD and a pI of 5. 2. Sequencing data from the N-terminus revealed the following initial 9 amino acids: (*)-Q-Q-(*)-E-L-Q-D-L.

6486311, Nov 26, 2002

Jun 29, 1998

09/106872

J. Steven Stanley - North Little Rock AR
Gael Cockrell - Cabot AR
Nina E. King - Little Rock AR
Hugh A. Sampson - Larchmont NY
Ricki M. Helm - LIttle Rock AR
Gary A. Bannon - LIttle Rock AR

Mt. Sinai School of Medicine - New York NY
University of Arkansas - Littlerock AR

C07H 2104

536 236, 4352523, 4241841

Peanuts are a common cause of food hypersensitivity reactions. The sera of 10 patients who had atopic dermatitis and a positive double-blind placebo-controlled food challenge to peanut were used to investigate the major allergens of peanut. Crude Florunner extracts were fractionated by anion-exchange chromatography using a step gradient (limit buffer, 0. 05M BisTris/1. 5M NaCl). A protein peak (OD 280) which eluted at 10% NaCl and demonstrated intense IgE-binding was further analyzed by two-dimensional SDS-PAGE/immunoblot analysis. The majority of this fraction is a protein which has a molecular weight of 17 kD and a pI of 5. 2. Sequencing data from the N-terminus revealed the following initial 9 amino acids: (*)-Q-Q-(*)-E-L-Q-D-L. Based on IgE-binding activity and no known amino acid sequence identity to other allergens, this allergen is designated Ara h II. Ara h II may be used to detect and quantify peanut allergens in foodstuffs.

6835824, Dec 28, 2004

Nov 13, 1998

09/191593

J. Steven Stanley - Little Rock AR
Gary A. Bannon - Little Rock AR
Gael Cockrell - Cabot AR
Ricki M. Helm - Little Rock AR

University of Arkansas - Little Rock AR

C07H 2104

536 236, 536 2532, 4242751, 4353201

One of the major peanut allergens, Ara h I, was selected from cDNA expression library clones using Ara h I specific oligo-nucleotides and polymerase chain reaction technology. The Ara h I clone identified a 2. 3 kb mRNA species on a Northern blot containing peanut poly A+RNA. DNA sequence analysis of the cloned inserts revealed that the Ara h I allergen has significant homology with the vicilin seed storage protein family found in most higher plants. The isolation of the Ara h I clones allowed the synthesis of this protein in cells and subsequent recognition of this. recombinant protein in immunoblot analysis using serum IgE from patients with peanut hypersensitivity.

7485708, Feb 3, 2009

Aug 26, 2002

10/228806

Gary A. Bannon - Wentzville MO, US
Hugh A. Sampson - Larchmont NY, US
Ricki M. Helm - Little Rock AR, US
J. Steven Stanley - North Little Rock AR, US
Patrick A. Rabjohn - Little Rock AR, US

University of Arkansas - Little Rock AR

C07H 21/04

536 231, 435325, 530370, 530806, 530868

It has been determined that allergens, which are characterized by both humoral (IgE) and cellular (T cell) binding sites, can be modified to be less allergenic by modifying the IgE binding sites. The IgE binding sites can be converted to non-IgE binding sites by masking the site with a compound that prevents IgE binding or by altering as little as a single amino acid within the protein, most typically a hydrophobic residue towards the center of the IgE binding epitope, to eliminate IgE binding. The method allows the protein to be altered as minimally as possible, other than within the IgE-binding sites, while retaining the ability of the protein to activate T cells, and, in some embodiments by not significantly altering or decreasing IgG binding capacity. The examples use peanut allergens to demonstrate alteration of IgE binding sites. The critical amino acids within each of the IgE binding epitopes of the peanut protein that are important to immunoglobulin binding have been determined.

7879977, Feb 1, 2011

Jan 10, 2006

11/329924

Gary A. Bannon - Wentzville MO, US
Hugh A. Sampson - Greenwich CT, US
Ricki M. Helm - Little Rock AR, US
Gael Cockrell - Cabot AR, US
J. Steven Stanley - North Little Rock AR, US
Nina E. King - Mason OH, US

University of Arkansas - Little Rock AK
Mount Sinai School of Medicine of New York University - New York NY

C07K 14/415
A61K 39/35

530350, 4241851, 4242751

It has been determined that allergens, which are characterized by both humoral (IgE) and cellular (T cell) binding sites, can be modified to be less allergenic by modifying the IgE binding sites. The IgE binding sites can be converted to non-IgE binding sites by masking the site with a compound that prevents IgE binding or by altering as little as a single amino acid within the protein, most typically a hydrophobic residue towards the center of the IgE-binding epitope, to eliminate IgE binding. The method allows the protein to be altered as minimally as possible, other than-within the IgE-binding sites, while retaining the ability of the protein to activate T cells, and, in some embodiments by not significantly altering or decreasing IgG binding capacity The examples use peanut allergens to demonstrate alteration of IgE binding sites. The critical amino acids within each of the IgE binding epitopes of the peanut protein that are important to immunoglobulin binding have been determined. Substitution of even a single amino acid within each of the epitopes led to loss of IgE binding.

20030202980, Oct 30, 2003

Mar 18, 2002

10/100303

Michael Caplan - Woodbridge CT, US
Howard Sosin - Fairfield CT, US
Hugh Sampson - Larchmont NY, US
Gary Bannon - Wentzville MO, US
A. Burks - Little Rock AR, US
Gael Cockrell - Cabot AR, US
Cesar Compadre - Little Rock AR, US
Cathie Connaughton - Conway AR, US
Ricki Helm - Little Rock AR, US
Nina King - Mason OH, US
Randall Kopper - Conway AR, US
Soheila Maleki - New Orleans LA, US
Patrick Rabjohn - Little Rock AR, US
David Shin - San Diego CA, US
J. Stanley - North Little Rock AR, US

A61K039/00
C07H021/04
C12P021/02
C12N005/06
C07K014/415
C07K014/47

424/185100, 435/069100, 435/320100, 435/325000, 530/350000, 530/370000, 536/023500, 536/023600

It has been determined that allergens, which are characterized by both humoral (IgE) and cellular (T-cell) binding sites, can be modified to be less allergenic by modifying the IgE binding sites. The IgE binding sites can be converted to non-IgE binding sites by altering as little as a single amino acid within the protein, preferably a hydrophobic residue towards the center of the IgE epitope, to eliminate IgE binding. Additionally or alternatively a modified allergen with reduced IgE binding may be prepared by disrupting one or more of the disulfide bonds that are present in the natural allergen. The disulfide bonds may be disrupted chemically, e.g., by reduction and alkylation or by mutating one or more cysteine residues present in the primary amino acid sequence of the natural allergen. In certain embodiments, modified allergens are prepared by both altering one or more linear IgE eitopes and disrupting one or more disulfide bonds of the natural allergen. In certain embodiments, the methods of the present invention allow allergens to be modified while retaining the ability of the protein to activate T-cells, and, in some embodiments by not significantly altering or decreasing IgG binding capacity. The Examples provided herein use peanut allergens to illustrate applications of the invention.

20120283421, Nov 8, 2012

Jul 15, 2011

13/183892

Michael J. Caplan - Woodbridge CT, US
Howard B. Sosin - Fairfield CT, US
Hugh A. Sampson - Greenwich CT, US
Gary A. Bannon - Wentzville MO, US
Gael Cockrell - Cabot AR, US
Cesar M. Compadre - Little Rock AR, US
Cathie Connaughton - Conway AR, US
Ricki M. Helm - Little Rock AR, US
Nina E. King - Mason OH, US
Randall A. Kopper - Conway AR, US
Soheila J. Maleki - New Orleans LA, US
Patrick A. Rabjohn - Little Rock AR, US
David S. Shin - San Diego CA, US
J. Steven Stanley - North Little Rock AR, US

C07K 14/00

530403, 530350

It has been determined that allergens, which are characterized by both humoral (IgE) and cellular (T-cell) binding sites, can be modified to be less allergenic by modifying the IgE binding sites. The IgE binding sites can be converted to non-IgE binding sites by altering as little as a single amino acid within the protein, preferably a hydrophobic residue towards the center of the IgE epitope, to eliminate IgE binding. Additionally or alternatively a modified allergen with reduced IgE binding may be prepared by disrupting one or more of the disulfide bonds that are present in the natural allergen. The disulfide bonds may be disrupted chemically, e.g., by reduction and alkylation or by mutating one or more cysteine residues present in the primary amino acid sequence of the natural allergen. In certain embodiments, modified allergens are prepared by both altering one or more linear IgE epitopes and disrupting one or more disulfide bonds of the natural allergen. In certain embodiments, the methods of the present invention allow allergens to be modified while retaining the ability of the protein to activate T-cells, and, in some embodiments by not significantly altering or decreasing IgG binding capacity. The Examples provided herein use peanut allergens to illustrate applications of the invention.

59731214, Oct 26, 1999

Mar 4, 1996

8/610424

A. Wesley Burks - Little Rock AR
Ricki M. Helm - Little Rock AR

University of Arkansas - Little Rock AR

C07K 14415

530370

Peanuts are a common cause of food hypersensitivity reactions. The sera of 10 patients who had atopic dermatitis and a positive double-blind placebo-controlled food challenge to peanut were used to investigate the major allergens of peanut. Crude Florunner extracts were fractionated by anion-exchange chromatography using a step gradient (limit buffer, 0. 05M BisTris/1. 5M NaCl). One hundred microliters of each 2. 0 ml fraction was dot-blotted onto nitrocellulose paper and IgE-binding activity assessed using the serum pool to select allergen-containing fractions. A protein peak (OD 280) which eluted at 10% NaCl and demonstrated intense IgE-binding was further analyzed by two-dimensional SDS-PAGE/immunoblot analysis. The majority of this fraction is a protein which has a molecular weight of 17 kD and a pI of 5. 2. Sequencing data from the N-terminus revealed the following initial 9 amino acids: (*)-Q-Q-(*)-E-L-Q-D-L.