Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Reexamination Certificate
1998-04-15
2003-11-04
Achutamurthy, Ponnathapu (Department: 1652)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
C435S023000, C435S471000
Reexamination Certificate
active
06642011
ABSTRACT:
BACKGROUND OF THE INVENTION
A. Field of the Invention
The present invention relates to a human protein sequence which can be used in several applications. Specifically, the novel human protein sequence can be used to design proteins which produce lower allergenic response in humans exposed to such proteins through the use of a predictive assay.
B. State of the Art
Serine proteases are a subgroup of carbonyl hydrolases. They comprise a diverse class of enzymes having a wide range of specificities and biological functions. Stroud, R.
Sci. Amer.,
131:74-88. Despite their functional diversity, the catalytic machinery of serine proteases has been approached by at least two genetically distinct families of enzymes: the subtilisins and the mammalian chymotrypsin related and homologous bacterial serine proteases (e.g., trypsin and
S. gresius
trypsin). These two families of serine proteases show remarkably similar mechanisms of catalysis. Kraut, J. (1977),
Ann. Rev. Biochem.,
46:331-358. Furthermore, although the primary structure is unrelated, the tertiary structure of these two enzyme families bring together a conserved catalytic triad of amino acids consisting of serine, histidine and aspartate.
Subtilisin is a serine endoprotease (MW 27,500) which is secreted in large amounts from a wide variety of Bacillus species and other microorganisms. The protein sequence of subtilisin has been determined from at least four different species of Bacillus. Markland, F. S., et al. (1983),
Honne
-
Seyler's Z. Physiol. Chem.,
364:1537-1540. The three-dimensional crystallographic structure of
Bacillus amyloliquefaciens
subtilisin to 2.5A resolution has also been reported. Wright, C. S., et al. (1969),
Nature,
221:235-242; Drenth, J., et al. (1972),
Eur. J. Biochem.,
26:177-181. These studies indicate that although subtilisin is genetically unrelated to the mammalian chymotrypsin like serine proteases, it has a similar active site structure. The x-ray crystal structures of subtilisin containing covalently bound peptide inhibitors (Robertus, J. D., et al. (1972),
Biochemistry,
11:2439-2449) or product complexes (Robertus, J. D., et al. (1976),
J. Biol. Chem.,
251:1097-1103) have also provided information regarding the active site and putative substrate binding cleft of subtilisin. In addition, a large number of kinetic and chemical modification studies have been reported for subtilisin (Philipp, M., et al. (1983),
Mol. Cell. Biochem.,
51:5-32; Svendsen, B. (1976),
Carlsberg Res. Comm.,
41:237-291; Markland, F. S.
Id.
) as well as at least one report wherein the side chain of methionine at residue 222 of subtilisin was converted by hydrogen peroxide to methionine-sulfoxide (Stauffer, D. C., et al. (1965),
J. Biol. Chem.,
244:5333-5338) and the side chain of serine at residue 221 converted to cysteine by chemical modification (Polgar, et al. (1981),
Biochimica et Biophysica Acta,
667:351-354.)
Proteins bearing some resemblance and/or homology to bacterial subtilisin have also been detected in humans as well (see e.g., Keifer et al.,
DNA and Cell Biol.,
Vol. 10, No. 10, pp. 757-769 (1991); Smeekens et al.,
J. Biol. Chem.,
Vol. 265, No. 6, pp. 2997-3000 (1990); Tomkinson et al.,
Biochem., Vol.
30, pp. 168-174 (1991)).
U.S. Pat. No. 4,760,025 (RE 34,606) discloses the modification of subtilisin amino acid residues corresponding to positions in Bacillus amyloliquefaciens subtilisin tyrosine −1, aspartate +32, asparagine +155, tyrosine +104, methionine +222, glycine +166, histidine +64, glycine +169, phenylalanine +189, serine +33, serine +221, tyrosine +217, glutamate +156 and alanine +152. U.S. Pat. No. 5,182,204 discloses the modification of the amino acid +224 residue in
Bacillus amyloliquefaciens
subtilisin and equivalent positions in other subtilisins which may be modified by way of substitution, insertion or deletion and which may be combined with modifications to the residues identified in U.S. Pat. No. 4,760,025 (RE 34,606) to form useful subtilisin mutants or variants. U.S. Pat. No. 5,155,033 discloses similar mutant subtilisins having a modification at an equivalent position to +225 of
B. amyloliquefaciens
subtilisin. U.S. Pat. Nos. 5,185,258 and 5,204,015 disclose mutant subtilisins having a modification at positions +123 and/or +274. U.S. Pat. No. 5,182,204 discloses the modification of many amino acid residues within subtilisin, including specifically +99, +101, +103, +107, +126, +128, +135, +197 and +204. U.S. Pat. No. 4,914,031 discloses certain subtilisin analogs, including a subtilisin modified at position +76.
Proteins, including proteases, used in industrial, pharmaceutical and commercial applications are of increasing prevalence. As a result, the increased exposure due to this prevalence has been responsible for some safety hazards caused by the sensitization of certain persons to those peptides, whereupon subsequent exposure causes extreme allergic reactions which can be injurious and even fatal. For example, proteases are known to cause dangerous hypersensitivity in some individuals. As a result, despite the usefulness of proteases in industry, e.g., in laundry detergents, cosmetics, textile treatment etc . . . , and the extensive research performed in the field to provide improved proteases which have, for example, more effective stain removal under detergency conditions, the use of proteases in industry has been problematic due to their ability to produce a hypersensitive allergenic response in some humans.
Much work has been done to alleviate these problems. Among the strategies explored to reduce immunogenic potential of protease use have been improved production processes which reduce potential contact by controlling and minimizing workplace concentrations of dust particles or aerosol carrying airborne protease, improved granulation processes which reduce the amount of dust or aerosol actually produced from the protease product, and improved recovery processes to reduce the level of potentially allergenic contaminants in the final product. However, efforts to reduce the allergenicity of protease, per se, have been relatively unsuccessful. Alternatively, efforts have been made to mask epitopes in protease which are recognized by immunoglobulin E (IgE) in hypersensitive individuals (PCT Publication No. WO 92/10755) or to enlarge or change the nature of the antigenic determinants by attaching polymers or peptides/proteins to the problematic protease.
When an adaptive immune response occurs in an exaggerated or inappropriate form, the individual experiencing the reaction is said to be hypersensitive. Hypersensitivity reactions are the result of normally beneficial immune responses acting inappropriately and sometimes cause inflammatory reactions and tissue damage. They can be provoked by many antigens; and the cause of a hypersensitivity reaction will vary from one individual to the next. Hypersensitivity does not normally manifest itself upon first contact with the antigen, but usually appears upon subsequent contact. One form of hypersensitivity occurs when an IgE response is directed against innocuous environmental antigens, such as pollen, dust-mites or animal dander. The resulting release of pharmacological mediators by IgE-sensitized mast cells produces an acute inflammatory reaction with symptoms such as asthma or rhinitis.
Nonetheless, a strategy comprising modifying the IgE sites will not generally be successful in preventing the cause of the initial sensitization reaction. Accordingly, such strategies, while perhaps neutralizing or reducing the severity of the subsequent hypersensitivity reaction, will not reduce the number or persons actually sensitized. For example, when a person is known to be hypersensitive to a certain antigen, the general, and only safe, manner of dealing with such a situation is to isolate the hypersensitive person from the antigen as completely as possible. Indeed, any
Achutamurthy Ponnathapu
Genencor International Inc.
Genencor International Inc.
Moore William W.
LandOfFree
Human protease and use of such protease for pharmaceutical... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Human protease and use of such protease for pharmaceutical..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Human protease and use of such protease for pharmaceutical... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3175238