Cleaning composition

Abstract

The present invention provides a composition comprising one or more protease enzymes and one or more of: a non-aromatic C1 to C6 sulfonic acid; a non-aromatic C1 to C6 sulfonate salt; a non-aromatic C1 to C6 ester of sulfuric acid; and a salt of a non-aromatic C1 to C6 ester of sulfuric acid. The present invention further provides a method of treating a surface comprising contacting the surface with a diluted composition described herein.

Description

CLEANING COMPOSITIONField of the Invention

[0001] This invention relates to compositions and methods for inactivating proteins, for example P-sheet amyloid proteins or prions, which can be misfolded proteins, and a means for treating and / or cleaning and / or disinfecting materials contaminated by proteins, for example P-sheet amyloid proteins or prions or by similar conformationally altered proteins.

[0002] Although the present invention will be described hereinafter with reference to its preferred embodiment, it will be appreciated by those of skill in the art that the spirit and scope of the invention may be embodied in many other forms.Background of the Invention

[0003] Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field.

[0004] Historically, infectious agents such as bacteria, fungi, parasites, and viruses have well established methods of control that involve various forms of disinfection and sterilization (e.g. steam sterilization, dry sterilization, pasteurization, sterile filtration, treatment with ethylene oxide, gluteraldehyde, phenols or other disinfecting chemicals, radiation, etc.). With viruses, there are also established methods for example lowering the pH to 4.0 or below, heating at 60 °C for extended periods, or use of organic solvents in high concentrations. In addition, UV treatment, formaldehyde and specific antiviral agents have been employed.

[0005] For some years now, new and previously unknown species of pathogenic agents have appeared and have been reported in scientific publications. These have been referred to as prions and present one of the greatest challenges facing the health care industry today. Prions are infectious proteins that differ from bacteria and other previously known infectious agents. A number of neurological diseases have been identified recently both in humans and animals, that appear to be attributable to prions. As detailed in PCT / US00 / 14353 (the content of which is incorporated herein by reference), human diseases attributed to prions include Kuru, Creutzfeldt-Jakob disease (CJD), Gerstmann-Straussler-Scheinker disease (GSS), and Fatal Familial Insomnia.(FFI).

[0006] In addition to prion diseases of humans, disorders of animals are included in the group of known prion diseases. Scrapie of sheep and goats is perhaps the most studied animal prion disease. Several lines of inquiry have suggested a link between variant CJD and apreceding epidemic of bovine spongiform encephalopathy (BSE). No successful therapeutic treatments have been developed and as a result these diseases are always fatal. Adding to the problem is the fact that the incubation period can be up to 30 years in humans and this factor presents a major challenge to the scientists involved, with some predicting an epidemic "in the pipeline".

[0007] Groups possibly at risk of infection include patients who may come into contact with infected medical instruments during surgery including dentistry, medical staff dissecting infected material, and healthcare workers responsible for cleaning and sterilizing instruments. There are also concerns that groups at risk may be broadened to include veterinarians, abattoir workers, butchers in contact with cows or beef primarily in Europe and more recently persons receiving blood transfusions or organs from donors incubating a prion disease.

[0008] The structure of prions has been the subject of intense investigation and different points of view have been expressed. Some scientists believe they are extremely small viruses, while most experts now believe that prions are actually infectious proteins without a DNA or RNA core. More particularly the consensus now is that the PrP gene of mammals expresses a protein which can be the soluble, non-disease, cellular form PrPc or can be an insoluble disease form PrPsc. Many lines of evidence indicate that prion diseases result from the transformation of the normal cellular form into the abnormal PrPSc form. There is no detectable difference in the amino acid sequence of the two forms. The PrPc form is composed of a highly membrane associated 33 - 35kDa protein which degrades on digestion with protease K. However the PrPSc form has an altered conformational form, in particular having a high level of B-sheet conformation. Properties of PrPSc useful in diagnosing the infective altered conformational form are a protease resistant core of 27 - 30kDa. Another distinctive feature of the altered conformational infective form is that it acquires a hydrophobic core.

[0009] Conventional disinfection and sterilizing agents have no significant effect on prions in an acceptable time. Attempts to deactivate prions and / or to disinfect surfaces on which they may be transmitted have shown an extraordinary resistance. The conditions required are generally too severe to be practical for routine disinfection, not only in terms of time and cost, but also in terms of damage to materials and occupational health hazards involved. For example in one study viable prion particles have been detected in a sample after 5-15mins / 600 °C dry heat although total destruction could be achieved at 1000 °C in 15mins and in 1- lOhrs at >200 °C. It has been proposed to treat with I M. caustic soda (pH14) for 2hrs butthat treatment is extremely corrosive, dangerous to staff, and aggressive to materials and has been shown not to eliminate the total Ml 000 prion load in a reasonable time period. US5633349 describes a procedure for treating a biological material involving treatment with 6-8 molar urea or 1-2 molar sodium thiocyanate for a minimum of 12 hrs (preferably 18 hrs) which suffers from similar disadvantages.

[0010] Because of the difficulties in decontamination it has been proposed as preferably that surgical instruments used in brain surgery should be used only once, but this implies a disposal risk in addition to being expensive and for some instruments impractical.PCT / US00 / 14353 describes a method of rendering prions non-infectious by use of a polycationic dendrimer but it is not clear whether that process is reversible or permanent or commercially viable for disinfecting surfaces.

[0011] Although attention has been focused on the PrPc form and the PrPSc form it has also been suggested that the protein can exist in an intermediate form which has a B-sheet content intermediate between the predominantly alpha helix structure of the PrPc form and the predominantly B-sheet conformation of the PrPSc form and which retains solubility in the absence of a denaturant.

[0012] The assembly or misassembly of normally soluble proteins into conformationally altered insoluble proteins is thought to be causative of, or implicated in, a variety of other diseases. For example, a-synuclein and tau are amyloid proteins which are strongly implicated in potential prion-like activity with respect to neurological diseases. Although the invention will be herein described in relation to P amyloid proteins or prions, it will be understood to be applicable to other insoluble or enzyme resistant conformationally altered proteins implicated in disease.

[0013] PCT / AU2002 / 00092 (WO 2002 / 062400 Al) relates to a methods and compositions for treating a surface, suspension or solution contaminated with a PrPprion protein or a surrogate thereof. The methods and compositions employ a combination of one or more enzymes effective to cleave a prion protein to fragments having a non-infective molecular weight, and one or more agents selected to favour conformational unfolding of the PrPprion protein while not denaturing the one or more enzymes.

[0014] It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative.

[0015] An object of the invention is to provide improved, or at least alternative, means of disinfecting a surface infected with proteins, such as P amyloid proteins or prions. In certain preferred embodiments, the invention renders prions inactive more efficiently, that is to saymore effectively in a given time, or as effectively in a shorter time, than prior art methods. Certain highly preferred embodiments of the invention achieve better than a 4 log reduction in less than 60 mins at below 60 °C. In some embodiments the invention is also applicable to prions in situations other than on surfaces for example in suspension in a solid, liquid or gaseous medium or in biological systems and may have other in vitro or in vivo uses.

[0016] Amyloid refers to the abnormal fibrous, extracellular, proteinaceous deposits found in organs and tissues. Amyloid is insoluble and is structurally dominated by P-sheet structure of the constituent amyloid proteins. Unlike other fibrous proteins it does not commonly have a structural, supportive or motility role but is associated with the pathology seen in a range of diseases broadly described as amyloidosis. These diseases include Alzheimer’s, the spongiform encephalopathies and type II diabetes, all of which are progressive disorders with associated high morbidity and mortality.

[0017] Amyloid fibrils are formed by normally soluble proteins, which assemble to form insoluble fibers that are resistant to degradation. Their formation can accompany disease and each disease is characterised by a specific protein or peptide that aggregates. Well known examples of amyloid diseases include Alzheimer's disease, Parkinsons disease, Diabetes type 2 and the spongiform encephalopathies (e.g., Mad cow disease). The amyloid fibrils are deposited extracellularly in the tissues and are thought to have a pathogenic effect. The fibrillar assemblies are inherently stable and structural studies have revealed that they are composed predominantly of P-sheet structure in a characteristic cross-P conformation.

[0018] The term "prion protein" as herein used includes variants, fragments, fusions, and analogues that have other interactions or activities that are substantially the same as those of a full length prion protein or amyloid protein sequence, but which may be more convenient to use and includes all forms of secondary structure The term is also herein used to include prion or amyloid surrogates, that is to say proteins which are not themselves prions / amyloids but which have similar structure or exhibit similar behaviour to prions or amyloids and can be used to model or predict how a prion or amyloid would perform under specified conditions. The term "PrPScprion protein" is intended to have a similarly broad meaning but is limited to prion proteins or amyloid proteins which by virtue of their secondary or tertiary structure are enzyme resistant and includes conformations which are similarly enzyme resistant.

[0019] Although the invention will be described with reference to specific examples it will be appreciated by those skilled in the art that the invention may be embodied in many other forms.Summary of the Invention

[0020] According to a first aspect, the invention provides a composition comprising one or more enzymes and one or more of:

[0021] a non-aromatic Cl to C6 sulfonic acid;

[0022] a non-aromatic Cl to C6 sulfonate salt;

[0023] a non-aromatic Cl to C6 ester of sulfuric acid; and

[0024] a salt of a non-aromatic Cl to C6 ester of sulfuric acid.

[0025] In a second aspect, the invention provides a composition comprising one or more protease enzymes,

[0026] a hydrotrope selected from the group consisting of water soluble hydrotropes having anions of formula:

[0027] wherein R1 and R2 are independently H or linear or branched Cl to C12 alkyl groups,

[0028] and one or more of:

[0029] a non-aromatic Cl to C6 sulfonic acid;

[0030] a non-aromatic Cl to C6 sulfonate salt;

[0031] a non-aromatic Cl to C6 ester of sulfuric acid; and

[0032] a salt of a non-aromatic Cl to C6 ester of sulfuric acid.

[0033] According to a third aspect, the invention provides a diluted composition comprising the composition of the first or second aspect and water in a ratio of compositiomwater of from 1 :2000 to 1 : 10.

[0034] According to a fourth aspect, the invention provides a method of treating a surface comprising contacting the surface with a diluted composition of the third aspect.

[0035] According to a fifth aspect, the invention provides a method of treating a suspension or solution comprising contacting suspension or solution with a diluted composition of the third aspect.Brief Description of the Drawings

[0036] Preferred embodiments of the invention will now be described, by way of example only, with reference to the accompanying drawings in which:

[0037] Figure 1 : Autozyme PR Sample [Al 12068], Autozyme PR Sample 1 (New Chaotrophic Agent) [B2373], and Autozyme PR (Prioncidal Challenge) [B2563] enzymatic digestion of PD alpha-synuclein TMR peak numbers with & without Control after digestion at 60 °C or 40 °C incubation for 10 minutes. Each variable had an enzymatic digestant concentration of 1 :100 concentration. Raw data from Table 1.

[0038] Figure 2: Autozyme PR Sample [Al 12068], Autozyme PR Sample 1 (New Chaotrophic Agent) [B2373], and Autozyme PR (Prioncidal Challenge) [B2563] enzymatic digestion of PD alpha-synuclein ThT peak numbers with & without Control after digestion at 60 °C or 40 °C incubation for 10 minutes. Each variable had an enzymatic digestant concentration of 1 :100 concentration. Raw data from Table 2.

[0039] Figure 3: Autozyme PR Sample [Al 12068], Autozyme PR Sample 1 (New Chaotrophic Agent) [B2373], and Autoyzme PR (Prioncidal Challenge) [B2563] enzymatic digestion of PD alpha-synuclein TMR Total Peak Intensity with & without Control after digestion at 60 °C or 40 °C incubation for 10 minutes. Each variable had an enzymatic digestant concentration of 1 : 100 concentration. Raw data from Table 3.

[0040] Figure 4: Autozyme PR Sample [Al 12068], Autozyme PR Sample 1 (New Chaotrophic Agent) [B2373], and Autoyzme PR (Prioncidal Challenge) [B2563] enzymatic digestion of PD alpha-synuclein ThT Total Peak Intensity with & without Control after digestion at 60 °C or 40 °C incubation for 10 minutes. Each variable had an enzymatic digestant concentration of 1 : 100 concentration. Raw data from Table 4.

[0041] Figure 5: Autozyme PR Sample [Al 12068], Autozyme PR Sample 1 (New Chaotrophic Agent) [B2373], and Autozyme PR (Prioncidal Challenge) [B2563] enzymatic digestion of PD alpha-synuclein TMR & ThT peak numbers with Control after digestion at 60 °C or 40 °C with incubation of 10 minutes. Each variable had an enzymatic digestant concentration of 1 : 100 concentration. Summarizing comparison between ThT and TMR Peak numbers.

[0042] Figure 6: Autozyme PR Sample [Al 12068], Autozyme PR Sample 1 (New Chaotrophic Agent) [B2373], and Autozyme PR (Prioncidal Challenge) [B2563] enzymatic digestion of PD a-synuclein TMR & ThT total peak intensity with Control after digestion at 60 °C or 40 °C with incubation of 10 minutes. Each variable had an enzymatic digestant concentration of 1 : 100 concentration. Summarizing comparison between ThT and TMR Peaknumbers.

[0043] Figure 7: Autozyme PR Sample [Al 12068], Autozyme PR Sample 1 (New Chaotrophic Agent) [B2373], and Autozyme PR (Prioncidal Challenge) [B2563] peak number control results for ThT interference without PD a-Synuclein with and without control peak numbers after digestion at 60 °C or 40 °C incubation for 20 minutes. Each variable had an enzymatic digestant concentration of 1 : 100 concentration. Raw data from Table 5.

[0044] Figure 8: Autozyme PR Sample [Al 12068], Autozyme PR Sample 1 (New Chaotrophic Agent) [B2373], and Autozyme PR (Prioncidal Challenge) [B2563] total peak intensity control results for ThT interference without PD a-Synuclein with and without control peak numbers after digestion at 60 °C or 40 °C incubation for 20 minutes. Each variable had an enzymatic digestant concentration of 1 : 100 concentration. Raw data from Table 6.Definitions

[0045] In describing and claiming the present invention, the following terminology will be used in accordance with the definitions set out below. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments of the invention only and is not intended to be limiting. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one having ordinary skill in the art to which the invention pertains.

[0046] Unless the context clearly requires otherwise, throughout the description and the claims, the words “comprise”, “comprising”, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”.

[0047] Throughout the description and the claims the expression “alkyl”, unless specifically limited, denotes a Cl -6 alkyl group, e.g. Cl -5 alkyl group, e.g. Cl -4 alkyl group, e.g. Cl -3 alkyl group, e.g. Cl-2 alkyl group, e.g. methyl. Alkyl groups are saturated and may be straight chain or branched. Suitable alkyl groups include, for example, methyl, ethyl, propyl (e.g. n-propyl and isopropyl), butyl (e.g n-butyl, iso-butyl, sec-butyl and tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n-hexyl).

[0048] The term “halogen” or “halo” comprises fluorine (F), chlorine (Cl) and bromine (Br).

[0049] Alkyl may optionally be substituted by one or more halo atoms (halogen substituents), e.g. one to three, e.g. one or two, e.g. one. Fluoroalkyl groups may compriseone or more fluoro substituents, e.g. one to three, e.g. one or two, e.g. one. Exemplary haloalkyl groups include fluoroalkyl e.g. CF3. Suitable haloalkyl groups include, for example, fluoromethyl, difluouom ethyl and trifluoromethyl.

[0050] Preferably, alkyl is unsubstituted.

[0051] More preferably, alkyl is unsubstituted ethyl (CH2CH3) or methyl (CH3), most preferably methyl (CH3).

[0052] Concentrations expressed as % w / w are based on the total weight of the composition (unless otherwise specified).

[0053] References to “the composition” refer to the composition before dilution with water. References to “the diluted composition” refer to the composition after dilution with water.

[0054] P is used as the Greek symbol “beta”.

[0055] The term “disinfecting” or “disinfection” means the destruction or inactivation of infectious agents, including bacteria, fungi, viruses and / or prions or other amyloid proteins.

[0056] Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein are to be understood as modified in all instances by the term “about”. The examples are not intended to limit the scope of the invention. In what follows, or where otherwise indicated, “%” will mean “weight %”, “ratio” will mean “weight ratio” and “parts” will mean “weight parts”.

[0057] The term ‘substantially’ as used herein shall mean comprising more than 50% by weight, where relevant, unless otherwise indicated.

[0058] The recitation of a numerical range using endpoints includes all numbers subsumed within that range (e.g., 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.80, 4, 5, etc.).

[0059] Further, unless expressly stated to the contrary, “or” refers to an inclusive or and not to an exclusive or. For example, a condition A or B is satisfied by any one of the following: A is true (or present) and B is false (or not present), A is false (or not present) and B is true (or present), and both A and B are true (or present).

[0060] The terms “preferred” and “preferably” refer to embodiments of the invention that may afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful and is not intended to exclude other embodiments from the scope of the invention.

[0061] It must also be noted that, as used in the specification and the appended claims, the singular forms “a”, “an” and “the” include plural referents unless the context clearly dictates otherwise.

[0062] As used herein, with reference to numbers in a range of numerals, the terms “about,” “approximately” and “substantially” are understood to refer to the range of -10% to +10% of the referenced number, preferably -5% to +5% of the referenced number, more preferably -1 % to +1 % of the referenced number, most preferably -0.1 % to +0.1 % of the referenced number. Moreover, with reference to numerical ranges, these terms should be construed as providing support for a claim directed to any number or subset of numbers in that range. For example, a disclosure of from 1 to 10 should be construed as supporting a range of from 1 to 8, from 3 to 7, from 1 to 9, from 3.6 to 4.6, from 3.5 to 9.9, from 8 to 10, and so forth.Description of a Preferred Embodiment

[0063] The invention provides a composition comprising one or more enzymes and one or more of:

[0064] a non-aromatic Cl to C6 sulfonic acid;

[0065] a non-aromatic Cl to C6 sulfonate salt;

[0066] a non-aromatic Cl to C6 ester of sulfuric acid; and

[0067] a salt of a non-aromatic Cl to C6 ester of sulfuric acid.

[0068] The composition of the present invention optionally comprises an enzymestabilizing system. The composition of the present invention optionally comprises a surfactant / hydrotrope system. The composition of the present invention optionally comprises other components. The composition of the present invention comprises water.

[0069] Compounds will be classified according to the primary function that they are believed to provide in the compositions of the present invention.

[0070] However, it will be appreciated that compounds that perform a primary function may also function in other capacities. For example, compounds described as hydrotropes, which can be present to improve the solubility of surfacants, need not function exclusively as hydrotropes, but may also provide a degree of surfactancy or may contribute to the denaturing of P-sheet amyloid proteins or prions.

[0071] Certain quaternary ammonium compounds added to impart biocidal activity may also contribute to surfactancy and thus could be labelled cationic surfactants. a) Denaturing agent

[0072] The composition of the present invention comprises one or more denaturing agents (e.g. from one to ten, e.g. one, two, three or four, e.g. one, two or three, e.g. one or two, e.g.one).

[0073] The one or more denaturing agents are selected from the list consisting of:• a non-aromatic Cl to C6 sulfonic acid;• a non-aromatic Cl to C6 sulfonate salt;• a non-aromatic Cl to C6 ester of sulfuric acid; and• a salt of a non-aromatic Cl to C6 ester of sulfuric acid.

[0074] Non-aromatic Cl to C6 is preferably Cl to C6 alkyl, is preferably Cl to C5 alkyl, more preferably Cl to C4 alkyl, more preferably Cl to C3 alkyl, e.g. Cl to C2 alkyl, e.g. methyl.

[0075] Alkyl may optionally be substituted by one or more halo atoms (e.g. fluoro), e.g. one to three, e.g. one or two, e.g. one (e.g. one fluoro atom). Alkyl is preferably unsubstituted.

[0076] The total concentration of denaturing agent in the composition is preferably in the range of from 1 to 20 % w / w, e.g. 2 to 15 % w / w, e.g. 5 to 10 % w / w, e.g. 6 to 8 % w / w, before dilution of the composition to afford a diluted composition. b) Sulfonic acid / sulfonate salt

[0077] In one embodiment the composition comprises a non-aromatic Cl to C6 sulfonic acid.

[0078] A non-aromatic Cl to C6 sulfonic acid can be represented by the formula

[0079] wherein Al is non-aromatic Cl to C6.

[0080] The non-aromatic Cl to C6 sulfonic acid is preferably a Cl to C6 alkyl sulfonic acid, e.g. a Cl to C5 alkyl sulfonic acid, e.g. a Cl to C4 alkyl sulfonic acid, e.g. a Cl to C3 alkyl sulfonic acid, a Cl to C2 alkyl sulfonic acid, e.g. methanesulfonic acid.

[0081] Cl to C6 alkyl is optionally substituted by one to three halo atoms (e.g. fluoro), e.g. one to two halo atoms, e.g. one halo atoms, e.g. one fluoro atom. Cl to C6 alkyl is preferably unsubstituted.

[0082] In one embodiment, the non-aromatic Cl to C6 sulfonic acid is methylsulfonic acid,

[0083] In one embodiment the composition comprises a non-aromatic Cl to C6 sulfonate salt.

[0084] An anion of a non-aromatic Cl to C6 sulfonate salt can be represented by the formulawherein A2 is non-aromatic Cl to C6. The anion may be referred to as a sulfonate anion.

[0085] The non-aromatic Cl to C6 sulfonate salt is preferably a Cl to C6 alkyl sulfonate salt, e.g. a Cl to C5 alkyl sulfonate salt, e.g. a Cl to C4 alkyl sulfonate salt, e.g. a Cl to C3 alkyl sulfonate salt, a Cl to C2 alkyl sulfonate salt, e.g. methyl sulfonate salt.

[0086] Cl to C6 alkyl is optionally substituted by one to three halo atoms (e.g. fluoro), e.g. one to two halo atoms, e.g. one halo atoms, e.g. one fluoro atom. Cl to C6 alkyl is preferably unsubstituted.

[0087] The non-aromatic Cl to C6 sulfonate salt cation may be any cation which forms a soluble salt, e.g. selected from the group consisting of sodium, potassium, magnesium and calcium. Preferably the cation is sodium or potassium, most preferably sodium.

[0088] Examplary sulfonate salts include sodium alkanesulfonate salts such as sodium methanesulfonate and sodium ethylsulfonate. A further example is sodium tri fluoromethanesulfonate.

[0089] In one embodiment, the non-aromatic Cl to C6 sulfonate salt is methyl sulfonate sodium salt,c) Sulfuric acid ester / salt of a sulfuric acid ester

[0090] In one embodiment the composition comprises a non-aromatic Cl to C6 ester ofsulfuric acid. The non-aromatic Cl to C6 ester of sulfuric acid is preferably a mono-ester.

[0091] A non-aromatic Cl to C6 mono-ester of sulfuric acid can be represented by the formula

[0092] wherein A3 is non-aromatic Cl to C6.

[0093] The non-aromatic Cl to C6 ester of sulfuric acid is preferably a Cl to C6 alkyl ester of sulfuric acid, e.g. a Cl to C5 alkyl ester of sulfuric acid, e.g. a Cl to C4 alkyl ester of sulfuric acid, e.g. a Cl to C3 alkyl ester of sulfuric acid, a Cl to C2 alkyl ester of sulfuric acid, e.g. mono-methyl ester of sulfuric acid (methyl sulfate, also known as methyl hydrogen sulfate, methyl sulfuric acid; monomethyl sulfate; metilsulfate).

[0094] Cl to C6 alkyl is optionally substituted by one to three halo atoms (e.g. fluoro), e.g. one to two halo atoms, e.g. one halo atoms, e.g. one fluoro atom. Cl to C6 alkyl is preferably unsubstituted.

[0095] In one embodiment, the non-aromatic Cl to C6 ester of sulfuric acid is methyl sulfate,

[0096] In one embodiment the composition comprises a salt of a non-aromatic Cl to C6 ester of sulfuric acid.

[0097] An anion of a salt of a non-aromatic Cl to C6 ester of sulfuric acid can be represented by the formula

[0098] wherein A4 is non-aromatic Cl to C6. The anion may be referred to as a sulfate anion.

[0099] The salt of a non-aromatic Cl to C6 ester of sulfuric acid is preferably the salt of a Cl to C6 alkyl ester of sulfuric acid, e.g. a salt of a Cl to C5 alkyl ester of sulfuric acid, e.g. a salt of a Cl to C4 alkyl ester of sulfuric acid, e.g. a salt of a Cl to C3 alkyl ester of sulfuricacid, a salt of a Cl to C2 alkyl ester of sulfuric acid, e.g. methyl sulfate sodium salt. [000100] Cl to C6 alkyl is optionally substituted by one to three halo atoms (e.g. fluoro), e.g. one to two halo atoms, e.g. one halo atoms, e.g. one fluoro atom. Cl to C6 alkyl is preferably unsubstituted.[000101] The cation of the non-aromatic Cl to C6 ester of sulfuric acid may be any cation which forms a soluble salt, e.g. selected from the group consisting of sodium, potassium, magnesium and calcium. Preferably the cation is sodium or potassium, most preferably sodium.[000102] Examplary salts include sodium salts of Cl to C6 alkyl esters of sulfuric acid such as sodium methanesulfate and sodium ethylsulfate. A further example is sodium tri fluoromethanesulfate.[000103] In one embodiment, the salt of a non-aromatic Cl to C6 ester of sulfuric acid is methyl sulfate sodium salt (also known as monomethyl ester sulfuric acid sodium salt and sodium methyl sulfate),d) Enzymes[000104] The composition of the invention comprises one or more protease enzymes (e.g. one, two, three or four, e.g. one, two or three, e.g. one or two, e.g. one).[000105] In one embodiment, the composition of the invention comprises one or more protease enzymes (e.g. one, two, three or four, e.g. one, two or three, e.g. one or two, e.g. one) and one or more other enzymes (e.g. one to nine, e.g. one, two, three or four, e.g. one, two or three, e.g. one or two, e.g. one), for example selected from groups such as: lipases, endocellulases, amylases, lysozymes, etc.[000106] The total concentration of the one or more enzymes in the composition is preferably in the range of 0.01 to 10% w / w, e.g. 0.01 to 1% w / w, e.g. 0.05 to 0.5% w / w, e.g.1 to 10% w / w, e.g. 5 to 8% w / w, e.g. 6 to 8% w / w, before dilution of the composition to afford a diluted composition. e) Enzyme-stabilising system[000107] The composition of the present invention optionally comprises an enzymestabilising system. The enzyme-stabilising system can comprise various components, forexample one or more of an inorganic salt, an organic solvent, a soluble boron compound and a formate compound. It can be understood that any new enzyme stabilizing systems that become known in the field can be applicable. j) Inorganic salt[000108] The enzyme-stabilising system optionally comprises one or more inorganic salts (e.g. one or two).[000109] Suitable inorganic salts include salts of 2+ metal cations, e.g. calcium and magnesium. g) Organic solvent[000110] The enzyme-stabilising system optionally comprises one or more organic glycols or glycol ether solvents (e.g. one, two, three or four, e.g. one, two or three, e.g. one or two, e.g. one).[000111] Suitable glycols include glycerol (also known as glycerin or glycerine) and propylene glycol (propane 1 ,2-diol). A further suitable glycol is ethylene glycol (ethane 1,2- diol).[000112] Suitable glycol ethers include methyl, ethyl, propyl and butyl ethers, e.g. methyl and ethyl ethers.[000113] An example of a suitable glycol ether is dipropylene glycol methyl ether (DPGME). Examples of glycol ethers also include polyethylene glycol, polypropylene glycol, and copolymers thereof.[000114] The total concentration of the one or more organic glycols or glycol ether solvents in the composition is preferably in the range of 2 to 10% w / w, e.g. 4 to 8% w / w, before dilution of the composition to afford a diluted composition. h) Soluble boron compound[000115] The enzyme-stabilising system optionally comprises one or more soluble boron compounds (e.g. one, two or three, e.g. one or two, e.g. one). “Soluble boron compound” refers to a boron compound that is soluble in water at room temperature.[000116] Examples of soluble boron compounds include sodium borate and boric acid. Anadditional example of a suitable soluble boron compound is phenylboronic acid. A further example of a suitable soluble boron compound is 4-Formylphenylboronic acid.[000117] The total concentration of the one or more soluble boron compounds in the composition is preferably in the range of 0 to 5% w / w, e.g. 0 to 4% w / w, e.g. 0 to 2% w / w, e.g. 2 to 4% w / w, e.g. 0.1 to 0.5% w / w, before dilution of the composition to afford a diluted composition. i) Formate[000118] The enzyme-stabilising system optionally comprises one or more formate compounds (e.g. one or two, e.g. one).[000119] An example of a suitable formate compound is sodium formate. Another example of a suitable formate compound is potassium formate.[000120] The total concentration of the one or more formate compounds in the composition is preferably in the range of 0 to 5% w / w, e.g. 0 to 3% w / w, e.g. 1 to 3% w / w, e.g. 0.1 to 2.5% w / w, before dilution of the composition to afford a diluted composition. j) Surfactant / hydrotrope system[000121] The composition of the present invention optionally comprises a surfactant / hydrotrope system. The surfactant / hydrotrope system may include one or more of a hydrotrope and may include one or more of a surfactant. k) Hydrotrope[000122] The surfactant / hydrotrope system optionally comprises one or more hydrotropes (e.g. one, two or three, e.g. one or two, e.g. one).[000123] A hydrotrope is a compound that solubilizes hydrophobic compounds in aqueous solutions.[000124] Suitably the hydrotrope is selected from the group consisting of water soluble hydrotropes having anions of formula:[000125] wherein R1and R2are independently H or linear or branched Cl to C 12 alkyl groups, preferably independently H or linear or branched Cl to C6 alkyl groups, preferably independently H or linear or branched Cl to C4 alkyl groups, more preferably independently H or linear or branched Cl to C3 alkyl groups.[000126] Preferably R1and R2are not both H. When R1 and / or R2 are alkyl, preferably alkyl is unsubstituted.[000127] In one embodiment, R1 is methyl and R2 is H. In another embodiment, R1and R2are methyl. In a further embodiment, R1 is isopropyl and R2 is H.[000128] Examples of water soluble hydrotropes include toluene sulfonate salt, xylene sulfonate salt and cumene sulfonate salt.[000129] Suitable counterions include sodium and potassium and other soluble counterions, preferably sodium. Thus, preferably the hydrotrope is selected from the group consisting of water soluble hydrotropes of the formula:[000130] Examples of water soluble hydrotropes include sodium toluene sulfonate, sodium xylene sulfonate and sodium cumene sulfonate.[000131] Other suitable hydrotropes include sodium-2-ethyl hexylsulfate, phosphate ester of oxyethylated phenol, amine alkylaryl sulfonate, naphthalene sulfonate, linear alkyl naphthalene sulfonate, sodium dihexyl sulfosuccinate, and sodium dodecylbenzene sulfonate. [000132] The total concentration of the one or more hydrotropes in the composition is preferably in the range of from 1 to 20 % w / w, e.g. 2 to 15 % w / w, e.g. 5 to 10 % w / w, e.g. 6 to 8 % w / w, before dilution of the composition to afford a diluted composition.I) Surfactant[000133] The surfactant / hydrotrope system optionally comprises one or more surfactants (e.g. one to five, e.g. one to four, e.g. one to three, e.g. one or two, e.g. one).[000134] Surfactants include anionic surfactants, cationic surfactants, amphoteric surfactants and nonionic surfactants.[000135] Anionic surfactants include compounds of the following types:(i) Linear and Branched Chain Alcohol Sulfates, Phosphates and Sulfonates(ii) Linear and Branched Chain ethoxylated Alcohol Sulfates, Phosphates and Sulfonates(iii) Alkyl Aryl Sulfates, Phosphates and Sulfonates(iv) Alkyl Aryl Ether Sulfates, Phosphates and Sulfonates(v) Alkali metal or Ammonium salts of Carboxylic acids eg Sodium Stearate(vi) Docusates eg Sodium dioctyl sulfosuccinate[000136] Cationic surfactants include compounds of the following types:(i) Alkyldimethylbenzyl Ammonium Chlorides(ii) Dialkyldimethyl Ammonium Chlorides(iii) Cetyltrimethylammonium bromide or chloride(iv) Cetylpyridiniumchloride or bromide(v) Benzethoniumchloride[000137] Amphoteric surfactants include compounds of the following types:(i) Sulfonates of primary, secondary or tertiary amines “Sultaines”(ii) Carboxylates of primary secondary or tertiary amines “Betaines”(iii) Tertiary amine oxides e.g. lauryl or myristyl dimethylamineoxide[000138] Nonionic surfactants include compounds of the following types:(i) Linear or branched chain alcohol ethoxylates(ii) Alkylphenol or Dialkylphenol ethoxylates(iii) Ethoxylated Fatty Acids eg polyethoxylated Tallow Amine(iv) Fatty acid Amides e.g. cocamide monoethanolamine or diethanolamine(v) Poloxamers (ethoxylated and / or propoxylated and / or butoxylated derivatives of branched or linear alcohols)(vi) Fatty acid esters e.g. glycerol monostearate or monoleate(vii) Fatty acid esters of sorbitol “SPAN” type(viii) Ethoxylated fattyacid estes of sorbitol “Tween” type(ix) Fatty acid esters of Sucrose(x) Alkylpolyglucosides (APGs)[000139] The total concentration of the one or more surfactants in the composition is preferably in the range of 0-30% w / w, before dilution of the composition to afford a diluted composition. m) Preservative[000140] Optionally the composition of the invention comprises one or more preservatives (e.g. one to five, e.g. one to four, e.g. one to three, e.g. one or two, e.g. one).[000141] Suitable preservatives are known to those skilled in the art.[000142] An example of suitable preservative is isothiazolin.[000143] Another example of a suitable preservative is phenoxyethanol.[000144] Another example of a suitable preservative is sodium pyridinethione.[000145] The total concentration of the one or more preservatives in the composition is preferably in the range of 0.0001 to 0.001% w / w, e.g. 0.0002 to 0.0006% w / w, before dilution of the composition to afford a diluted composition. n) Biocide[000146] Optionally the composition of the invention comprises one or more biocides (e.g. one, two or three, e.g. one or two, e.g. one).[000147] An example of a suitable biocide is a quaternary ammonium biocide (“qua ’) such as an aryl or alkyl quaternary ammonium compound, preferably a benzalkonium halide such as a benzalkonium chloride.[000148] Another example of a suitable biocide is dodecyl dimethyl ammonium carbonate.[000149] A further example of a suitable biocide is a biguanide (e.g. Polyhexamethylene biguanide (PHMB) or chlorhexidine gluconate).[000150] The total concentration of the one or more biocides in the composition is preferably in the range of 0-6% w / w, before dilution of the composition to afford a diluted composition. o) Colourant[000151] Optionally the composition of the invention comprises a colourant.[000152] Suitable colourants are known to those skilled in the art. An example is FD&C Blue No. l.[000153] The total concentration of the colorant in the composition is preferably in the range of 0 to 0.002 % w / w, e.g. 0.001% w / w, before dilution of the composition to afford a diluted composition. p) pH adjuster[000154] The pH of the composition can be adjusted for example using one or more bases (e.g. one, two or three, e.g. one or two, e.g. one). Suitable bases include, for example, sodium hydroxide and triethanolamine.[000155] The pH of the composition can be adjusted, for example, using one or more acids. Suitable acids include, for example, citric acid. q) H[000156] The pH of the composition is chosen with regard to the preferred characteristics of the components of the composition and / or with regard to the preferred end use of the composition. For example, a pH range may be chosen that will match the pH known to optimize the activity of one or more enzymes utilised in the composition. For example, for acidic enzymes, a pH in the range 4.5-7 may be preferred. For example, for alkaline enzymes, a pH in the range 7-10 may be preferred.[000157] The skilled person appreciates that the pH of the composition and the diluted composition may slightly differ as a consequence of the dilution with water. r) Water[000158] The composition of the present invention comprises water. Preferably, the composition of the present invention is an aqueous solution. s) Shelf-stability[000159] The present invention provides a composition which is preferably shelf-stable and can be stored at room temperature (22 °C) for at least 6 months, preferably at least 1 year.t) Dilution[000160] The present invention provides a composition which is intended for dilution with water to afford a diluted composition. The diluted composition is intended to be used in the method of the invention.[000161] Typically, the composition of the invention will be sold in the form of a concentrate and the composition is diluted to afford a diluted composition before use.[000162] When the composition is diluted with water, the ratio of compositiomwater is from 1 :2000 to 1 : 10, for example 1 : 1000 to 1 : 10, for example 1 :2000 to 1 :25, for example 1 : 1000 to 1 :25, for example 1 :750 to 1 :25, or 1 :500 to 1 :50, or 1 : 1000 to 1 :25.[000163] Thus, the diluted composition comprises the composition of the invention and water in a ratio of composition: water of from 1 :2000 to 1 : 10, for example 1 : 1000 to 1 : 10, for example 1 :2000 to 1 :25, for example 1 : 1000 to 1 :25, for example 1 :750 to 1 :25, or 1 :500 to 1 :50, or 1 : 1000 to 1 :25.Method[000164] The present invention provides a method of treating a surface comprising contacting the surface, suspension or solution with the diluted composition.[000165] In one embodiment, the method of treating is a method of cleaning or a method of disinfecting. Thus, the present invention provides a method of cleaning a surface comprising contacting the surface with the diluted composition. The present invention also provides a method of disinfecting a surface comprising contacting the surface with the diluted composition.[000166] The present invention provides a method of treating a suspension or solution comprising contacting the suspension or solution with the diluted composition.[000167] In one embodiment, the method further comprises the step of diluting a composition of the invention with water to afford a diluted composition.[000168] In one embodiment, the surface is the surface of a medical or dental or surgical or veterinary instrument or device.[000169] In one embodiment, the surface is a food preparation surface or a manufacturing surface or a hospital work surface.[000170] In the method of the present invention, the contact time is preferably at least one minute (e.g. about five minutes, e.g. about ten minutes, e.g. about fifteen minutes, e.g. abouttwenty minutes, about thirty minutes). “Contact time” refers to the contact time for the diluted composition with the surface to be cleaned.[000171] In the method of the present invention, the preferred contact temperature is dependent on the chosen enzyme(s). In one embodiment, the contact temperature is from 15 to 90 °C, e.g. from 20 to 60 °C, e.g. from 20 to 40 °C, e.g. from 30 to 50 °C.[000172] In one embodiment (e.g. when the surface to be cleaned is the surface of a medical or dental or veterinary or surgical instrument or device), the diluted composition of the present invention can be used in a stagnant bath / sink, an ultrasound bath, and automated washer or other similar equipment.[000173] The surface can optionally be rinsed with water after contact with the diluted composition of the present invention.[000174] In one embodiment, the treatment is selected so as to result after cleavage in a predetermined percentage of the protein fragments having a molecular weight of less than a predetermined molecular weight.[000175] In one embodiment of the method of the invention, at least 90% of the protein fragments after cleavage have a molecular weight of less than 27 kDa[000176] In one embodiment of the method of the invention, at least 90% of the protein fragments after cleavage have a molecular weight of less than 25 kDa.[000177] In one embodiment of the method of the invention, at least 90% of the protein fragments after cleavage have a molecular weight of less than 23 kDa.[000178] In one embodiment of the method of the invention, the conditions are selected to protect or refold the one or more enzymes while irreversibly unfolding or at least opening the prion sufficiently for access by the enzyme.[000179] In one embodiment, the method of the invention comprises application of one or more agents selected from the group consisting of irradiation, electric field, magnetic field, energetic vibration and combinations thereof.[000180] In one embodiment of the method of the invention, the energetic vibration is one or more of ultrasound, electromagnetic or mechanical vibration.[000181] In one embodiment, the method of the invention comprises application of one or more agents selected to promote unfolding from the group consisting of heat, pH, organic solvents of the kind which tend to denature proteins, chaotropic agents, surfactants tending to bind proteins, inorganic salts which are strong denaturants of proteins, agents which cause S — S bond scission, substances having a strong affinity for hydrophilic residues of amino acids, substances having a strong affinity for hydrophobic residues of amino acids,substances promoting adsorption on surfaces, anionic surfactants and combinations of the foregoing.[000182] In one embodiment, the one or more agents are selected to promote or protect folding of the one or more enzymes is selected from the group consisting of nucleophilic solvents, weakly protic stabilizing solvents, non ionic surfactants, ionic surfactants, zwitterionic and amphoteric surfactants, buffers, surface active homo-co- or block copolymers, sulfated compounds, deoxycholate, glycosaminoglycans and combinations of the foregoing.Conditions selected to favour conformation unfolding[000183] Ultrasonic energy, infrared and microwave radiation, high pressure, and subjecting protons to the action of electric and / or magnetic fields may be able to promote unfolding (refolding), and even shaking or stirring may be influential. These conditions may optionally be used in the method of the present invention.Examples[000184] The following compositions were prepared as detailed in Table A:[000185] The comparative formulation (Al 12068) comprises an aromatic sulfonate.[000186] Composition 1 (B2373) and composition 2 (B2563) are according to the invention and comprise an alkyl sulfonic acid (e.g. methanesulfonic acid).[000187] Alpha-Synuclein fibrils were tagged with a fluorescent agent that binds to fibrils (replicating stacks of synuclein monomer) but not to digested monomers that no longer replicate. Both the fluorescent dye and the fluorescent marker on the proteins were measured.[000188] There are two sets of graphs, one for peak intensity results and one for peak number results. Most of the graphs show the data both with and without control, to enable the difference between the current formulation and the new formulations to be more visible.Objective[000189] Testing of the enzymatic digestant agents:[000190] Autozyme PR Sample (Batch No. Al 12068)[000191] Autozyme PR Sample I “New Chaotrophic Agent” (Batch No. B2373)[000192] Autozyme PR- “Prion Challenge” (Batch No. B2563)[000193] Each active digestant was used in an enzymatic digestive reaction against seeded alpha-Synuclein TMR (Atto565 tagged) labelled as “PD” as these fibrils were incubated and were found to be most suited for accurate results in this experiment and derived from patient sample.Materials and methods[000194] A fibril concentration of a 1 : 1000 dilution of fibrils (alpha-Synuclein TMR + Phosphate Buffer Saline [PBS] + lOpM ThT) was previously found to be an appropriateconcentration to mimic real world scenarios while maximizing output from the fluorescent spectroscopy for this experiment.[000195] For the Control, to accurately compare the Autozyme’s enzymatic digestion, a 1 : 10,000 dilution was used then the final results (Peak number & Total Peak intensity) multiplied by lOx to imitate a 1 : 1000 dilution. The microscope images more precisely when fibrils are in a higher dilution thus explaining why the original concentration was used. The following components were combined at the following volumes:[000196] 1) [(1 pL of PD alpha-Synuclein) + (999 pL of PBS)] (1 : 1000)[000197] 2) [(4 pL of 1) + (4 pL of 100 pM ThT) + (32 pL of H2O)] (1 : 10)[000198] For the correct Autozyme PR Al 120681, Autozyme PR B23732, and Autozyme PR B25633 concentrations, each were initially diluted to a 1 : 10 concentration with MilliQ H2O. The enzymatic digestant concentrations were made with the following volumes: [000199] 1 : 10= [(100 pL Digestant 1,2,3 ) + (900 pL MilliQ H2O)]** This starting dilution would provide a final concentration of 1 : 100 final concentration in final incubated solution once combined with other components.[000200] Following this, to achieve the correct 1 : 1000 PD alpha-Synuclein Fibril concentration, the following method was followed:[000201] 1) [(1 pL PD alpha-Synuclein) + (99 pL PBS)] (1 : 100)[000202] Subsequently, for final digestive testing, the following mixtures was made:[000203] (4pL Autozymel,2,3 [1 : 10]) + (4 pL PD alpha-Synuclein [1 : 100]) + (4 pL 100 pM ThT) + (28 pL H2O)[000204] Each mixture was then incubated for either 10 minutes or 20 minutes at either 40 °C or 60 °C.[000205] As a further measure of control, it was deemed important to analyze the ThT Interference values (if any) to confirm if any of the obtained results were skewed by such interferences. To achieve this control, the same concentrations of Autozyme agents & ThT were used with the absence of any PD alpha-Synuclein.[000206] (4 pL Autozyme 1,2,3 [1 : 10]) + (4 pL 100 pM ThT) + (32 pL H2O)[000207] The results of this test are in Tables 5 and 6 and the accompanying Figures 7 and 8.Table 1: Summary of peak numbers from the TMR dye traceTable 2: Summary of peak numbers from the ThT dye trace* All three Anazome conditions obtained via fluorescent spectroscopyTable 3: Summary of total peak intensity from the TMR dye trace* All three Anazome conditions obtained via fluorescent spectroscopyTable 4 summarises the raw number, mean, and standard deviation of Total Peak Intensity from the ThT Dye trace of all three Autozyme conditions obtained via fluorescent spectroscopy.Table 4: Total peak intensity from ThT dye traceTable 5 summarises the raw number, mean, and standard deviation of Peak numbers from the ThT Dye trace of all three Autozyme conditions obtained via fluorescent spectroscopy to test ThT Interference.Table 5: Peak numbers from ThT dye traceTable 6 summarises the raw number, mean, and standard deviation of Total Peak Intensity from the ThT Dye trace of all three Autoclave conditions obtained via fluorescent spectroscopy to test ThT Interference.Table 6: Raw number, mean, and standard deviation of Total Peak Intensity from the ThT dye traceDiscussion[000208] The results obtained from the fluorescent spectroscopy analysis support the evidence that the each of the provided digestive agents positively initiate enzymatic digestion on PD alpha-Synuclein TMR tagged with Thioflavin T (ThT) to a certain extent. An overall reduction of peaks is observed in from the Control to each of the experimental conditions and reinforce the hypothesis of the induced enzymatic digestion. The overall peak intensities also decrease after the both the 40 °C and 60 °C, 10 or 20 minute incubations used across all conditions.[000209] In terms of which digestant is “most effective” at assimilating the PD alpha- Synuclein, the peak number and total peak intensity decrease percentages can be calculated. [000210] ([Peak Number / Control]*100 = x, Percent Decrease = 100-x)Table 7: TMR v ThT number and percentage decrease* < 90%; ** 90% < x < 95%; *** 95% < x[000211] There is an apparent increase in the overall ThT interference in the Autozyme PR 1 (New Chaotrophic agent) B2373 at 40 °C (Figure 7 & 8) which was also observed in the total peak intensity graphs of the experiments containing PD alpha-Synuclein in preceding figures and results. The results at 60 °C show a clear decrease in peak number and total peak intensity with respect to the comparative formulation.[000212] The Batch No. B2563has the highest overall percent decreases in peak number and total peak intensity in both TMR & ThT at both temperatures.[000213] Although the invention has been described with reference to specific examples it will be appreciated by those skilled in the art that the invention may be embodied in many other forms.

Claims

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:-1. A composition comprising one or more protease enzymes and one or more of: a non-aromatic Cl to C6 sulfonic acid; a non-aromatic Cl to C6 sulfonate salt; a non-aromatic Cl to C6 ester of sulfuric acid; and a salt of a non-aromatic Cl to C6 ester of sulfuric acid.

2. A composition according to claim 1, wherein the non-aromatic Cl to C6 sulfonic acid is a Cl to C6 alkyl sulfonic acid, preferably a Cl to C3 alkyl sulfonic acid.

3. A composition according to claim 1 or claim 2, wherein the non-aromatic Cl to C6 sulfonate salt is a Cl to C6 alkyl sulfonate salt, preferably a Cl to C3 alkyl sulfonate salt.

4. A composition according to any one of claims 1 to 3, wherein the non-aromatic Cl to C6 ester of sulfuric acid is a Cl to C6 alkyl ester of sulfuric acid, preferably a Cl to C3 alkyl ester of sulfuric acid.

5. A composition according to any one of claims 1 to 4, wherein the salt of a non- aromatic Cl to C6 ester of sulfuric acid is a salt of a Cl to C6 alkyl ester of sulfuric acid, preferably a salt of a Cl to C3 alkyl ester of sulfuric acid.

6. A composition according to any one of claims 1 to 5, wherein alkyl is unsubstituted.

7. A composition according to claim 1, wherein the non-aromatic Cl to C6 sulfonic acid is methanesulfonic acid.

8. A composition according to any one of claims 1 to 7, comprising a hydrotrope selected from the group consisting of water soluble hydrotropes having anions of formula:wherein R1and R2are independently H or linear or branched Cl to C 12 alkyl groups.

9. A diluted composition comprising the composition of any one of claims 1 to 8, and water in a ratio of composition: water of from 1 :2000 to 1 : 10.

10. A method of treating a surface comprising contacting the surface with a diluted composition of claim 9.

11. A method according to claim 10, further comprising diluting the composition of any one of claims 1 to 8 with water to afford the diluted composition of claim 9.

12. A method according to claim 10 or claim 11, wherein the surface is the surface of a medical or dental or veterinary or surgical instrument or device.

13. A method according to claim 10 or claim 11, wherein the surface is a food preparation surface or a manufacturing surface or a hospital work surface.

14. A method of treating a suspension or solution comprising contacting suspension or solution with the diluted composition according to claim 9.

15. A method according to claim 14, further comprising diluting the composition of any one of claims 1 to 8 with water to afford the diluted composition of claim 9.

16. A method according to any one of claims 10 to 15, wherein the treatment is selected so as to result after cleavage in a predetermined percentage of the protein fragments having a molecular weight of less than a predetermined molecular weight.

17. A method according to claim 16, wherein at least 90% of the protein fragments after cleavage have a molecular weight of less than 27 kDa.

18. A method according to claim 16, wherein at least 90% of the protein fragments after cleavage have a molecular weight of less than 25 kDa.

19. A method according to claim 16, wherein at least 90% of the protein fragments after cleavage have a molecular weight of less than 23 kDa.

20. A method according to any one of claims 16 to 19, wherein the conditions are selected to protect or refold the one or more enzymes while irreversibly unfolding or at least opening the prion sufficiently for access by the enzyme.

21. A method according to any one of claims 16 to 19, further comprising application of one or more agents selected from the group consisting of irradiation, electric field, magnetic field, energetic vibration and combinations thereof.

22. A method according to claim 21, wherein the energetic vibration is one or more of ultrasound, electromagnetic or mechanical vibration.

23. A method according to any one of claims 16 to 21, further comprising application of one or more agents selected to promote unfolding from the group consisting of heat, pH, organic solvents of the kind which tend to denature proteins, chaotropic agents, surfactants tending to bind proteins, inorganic salts which are strong denaturants of proteins, agents which cause S — S bond scission, substances having a strong affinity for hydrophilic residues of amino acids, substances having a strong affinity for hydrophobic residues of amino acids, substances promoting adsorption on surfaces, anionic surfactants and combinations of the foregoing.

24. A method according to claim 23, wherein the one or more agents selected to promote or protect folding of the one or more enzymes is selected from the group consisting of nucleophilic solvents, weakly protic stabilizing solvents, non ionic surfactants, ionic surfactants, zwitterionic and amphoteric surfactants, buffers, surface active homo-co- or block copolymers, sulfated compounds, deoxycholate, glycosaminoglycans and combinations of the foregoing.

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