Claims for Patent: 10,308,929

Summary for Patent: 10,308,929

Title:	Method of preparing biological material
Abstract:	This invention pertains to a method of preparing biological material from a biological sample selected from the group consisting of blood and sputum samples. The method includes the step of altering at least one constitutive characteristic of the biological sample in the presence of a capturing scaffold by adding a lysis buffer containing a solubilising agent and a detergent to the biological sample, for simultaneously inhibiting coagulation of the biological sample; lysing the biological sample to release the biological material from the biological sample, thus making the biological material available; and capturing at least one fraction of the biological material on the capturing scaffold.
Inventor(s):	Grobler; Anne Frederica (Potchefstroom, ZA), Levanets; Oksana (Potchefstroom, ZA)
Assignee:	North-West University (Potchefstroom, ZA)
Application Number:	14/343,019
Patent Claims:	1. A method of preparing biological material from a biological sample selected from the group consisting of blood and sputum samples, the method including the step of altering at least one constitutive characteristic of the biological sample in the presence of a capturing scaffold by adding a lysis buffer containing a solubilising agent in the form of a chaotropic salt selected from the group consisting of urea, thiourea, guanidine hydrochloride, lithium perchlorate, sodium iodine, sodium perchlorate, guanidine isothiocyanate, guanidine carbonate, guanidine thiocyanate, derivatives of said chaotropic salts and combinations thereof and a detergent to the biological sample, and concomitantly adding purified starch to the biological material together with the lysis buffer for neutralising any PCR inhibitors present in the biological material, and concomitantly physically treating the biological sample through agitation and elevating the temperature of the biological sample above 40 degrees Celsius and up to a 100 degrees Celsius; for simultaneously: inhibiting coagulation of the biological sample; lysing the biological sample to release biological material from the biological sample thus making the biological material available; and capturing at least one fraction of the biological material on the capturing scaffold in a single chamber or well such that the biological material captured on the capturing scaffold is usable directly for analysis without any purification required and wherein the biological material captured on the capturing scaffold can be subsequently air dried and stored at ambient temperature. 2. A method according to claim 1 including the subsequent step of removing the capturing scaffold together with the captured fraction of the biological sample from the remainder of the biological sample. 3. A method according to claim 1 wherein the concentration of the chaotropic salt in the lysis buffer is in the range of from 2 M to 8 M. 4. A method according to claim 1 wherein the lysis buffer is selected from the group consisting of phosphate buffers, 4-(2-hydroxyethyl)-1 -piperazineethanesulfonic acid (HEPES), N-Cyclohexyl-2-aminoethanesulfonic acid (CHES), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS) and piperazine-N/, N-bis(2-ethanesulfonic acid) (PIPES), Tris-HCI, as well as other tris(hydroxymethyl)aminomethane (Tris) buffers containing ethylene diamine tetra-acetic acid (EDTA), ethylene glycol tetra-acetic acid (EGTA), deoxycholate, sodium chloride (MCI), sodium phosphate, octylphenoxypolyethoxyethanol, and non-ionic surfactants provided with a hydrophilic polyethylene oxide group and a hydrocarbon lipophilic or hydrophobic group, and combinations thereof. 5. A method according to claim 4 wherein the lysis buffer has a pH of between 4 and 12. 6. A method according to claim 1 wherein the detergent is selected from the group consisting of 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS), a nonionic surfactant and emulsifier derived from polyethoxylated sorbitan and oleic acid, a nonionic surfactant which has a hydrophilic polyethylene oxide group and a hydrocarbon lipophilic or hydrophobic group, saponin, sodium deoxycholate, SDS, octyl glucoside, octyl thioglucoside, laurly maltose, octylphenoxypolyethoxyethanol, and combinations thereof. 7. A method according to claim 6 wherein the detergent has a concentration of between 0.3% and 6%. 8. A method according to claim 1 wherein the biological material is captured in the form of nucleic acids, protein, serum, cells, tissue, plasma, antigens, antibodies, or reaction products. 9. A method according to claim 1 including the step of concomitantly adding a reducing agent selected from the group consisting of 2-mecarptoethanol, dithiothreitol (DTT), 2- mercaptoethylamine, tris(2-carboxyl)phosphine (TCEP), cysteine HCI N-ethylmaleimide, Nacystelyn, dornase alfa, thymosin .beta.4, guaifenesin TCEP HCl, and combinations thereof, to the biological sample together with the lysis buffer containing the solubilising agent and the detergent. 10. A method according to claim 1 wherein the capturing scaffold is treated chemically and/or physically prior to the capturing of the biological material such that a batch of capturing scaffolds is pre-prepared and stored for later use. 11. A method according to claim 10 wherein the step of pre-treating the capturing scaffold physically includes the further step of increasing the outer surface area of the capturing scaffold. 12. A method according to claim 10 wherein the capturing scaffold is chemically pre-treated with a cross-linking agent selected from the group consisting of ethyldimethylaminopropyl carbodiimide (EDC), N,N'-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), 1-cyclohexyl-3-(2-morpholinyl-(4)-ethyl)carbodiitnide metho-p- toluenesulfonate (CMC), aldehyde and combinations thereof. 13. A method according to claim 10 wherein the step of pre-preparing the capturing scaffold chemically and/or physically includes the further steps of washing the treated capturing scaffold with Tris buffer (containing NaCI, a non-ionic surfactant and emulsifier derived from polyethoxylated sorbitan and oleic acid), de-ionised water, phosphate buffer containing a non-ionic surfactant emulsifier and combinations thereof. 14. A method according to claim 1 wherein the capturing scaffold is selected from the group consisting of nano- or micro-particles and a body of a polymeric material. 15. A method according to claim 14 wherein the nano-particles are prepared from polylactic acid or chitosan derivatives. 16. A method according to claim 14 wherein the polymeric material is selected from the group consisting of polyethylene, polystyrene, polypropylene, polyvinyl chloride, nylon, teflon (poly tetra polyethylene), polychloroprene and polyacrylonitrile.

Details for Patent 10,308,929

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	PULMOZYME	dornase alfa	Solution	103532	12/30/1993		2031-09-06