Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
65 words
A THF active composition having a molecular weight of 3323±220; an isoelectric point of from 5.7 to 5.9; an amino acid composition of approximately: aspartic acid--4 units; threonine--1 unit; serine--5 units; glutamic acid--8 units; proline--2 units; glycine--5 units; alanine--2 units; leucine--1 unit; lysine--1 unit; arginine--2 units; 1/2 cystine--1 to 2 units; being the product obtainable by purification of thymus tissue and removal therefrom of endotoxins.
Claim 2
Dependent Claim
65 words
A THF active composition having a molecular weight of 3323±220; an isoelectric point of from 5.7 to 5.9; an amino acid composition of approximately: aspartic acid--4 units; threonine--1 unit; serine--5 units; glutamic acid--8 units; proline--2 units; glycine--5 units; alanine--2 units; leucine--1 unit; lysine--1 unit; arginine--2 units; 1/2 cystine--1 to 2 units; being the product obtainable by purification of thymus tissue and removal therefrom of endotoxins.
Claim 3
Dependent Claim
114 words
A process for preparing substantially pure and endotoxin-free THF which comprises: (a) comminuting fresh thymus tissue from mammals in a suitable liquid medium; at a pH between 7 and 8; (b) removing cell debris and other cell residues by centrifugation and ultracentrifugation; (c) separating the supernatant liquid and filtering same through suitable filtration media to remove particulate material and so as to sterilize the passing liquid and to remove endotoxins, (d) subjecting the liquid to exhaustive dialysis; (e) lyophilizing the dialyzate and redissolving it in a suitable liquid medium; (f) subjecting the lyophilized redissolved material to gel filtration and assaying the resulting peaks (fractions), and (g) eluting the active constitutent in substantially pure form.
Claim 4
Dependent Claim
114 words
A process for preparing substantially pure and endotoxin-free THF which comprises: (a) comminuting fresh thymus tissue from mammals in a suitable liquid medium; at a pH between 7 and 8; (b) removing cell debris and other cell residues by centrifugation and ultracentrifugation; (c) separating the supernatant liquid and filtering same through suitable filtration media to remove particulate material and so as to sterilize the passing liquid and to remove endotoxins, (d) subjecting the liquid to exhaustive dialysis; (e) lyophilizing the dialyzate and redissolving it in a suitable liquid medium; (f) subjecting the lyophilized redissolved material to gel filtration and assaying the resulting peaks (fractions), and (g) eluting the active constitutent in substantially pure form.
Claim 5
Dependent Claim
35 words
A process according to claim 2, wherein the thymus is comminuted in 1 to 5 volumes of phosphate buffer, in saline, tris-buffer or the like, at a pH of from pH 7 to pH 8.
Claim 6
Dependent Claim
24 words
A process according to claim 2, wherein after ultracentrifugation, the remaining liquid is filtered through membrane filters of from 0.8 to about 0.45 μm.
Claim 7
Dependent Claim
30 words
A process according to claim 2, wherein the liquid obtained after removal of particulate matter is subjected to exhaustive dialysis against larger volumes of water, saline, PBS or the like.
Claim 8
Dependent Claim
38 words
A process according to claim 2, wherein the lyophilized dialysate is redissolved in distilled water, in ammonium bicarbonate, PBS, tris-buffer or the like so as to obtain a protein concentration of about 1 to 5 mg/ml of solvent.
Claim 9
Dependent Claim
20 words
A process according to claim 2, wherein the gel filtration is effected on Sephadex columns ranging from G-10 to G-50.
Claim 10
Dependent Claim
24 words
A process according to claim 2, wherein the fractions eluted by ion exchange chromatography are obtained from peaks substantially as defined by FIG. 1A.
Claim 11
Dependent Claim
37 words
A THF active composition obtainable by extraction from thymus cells by collection of the component eluted at 0.15 M NaCl from an anion exchange chromatograph on DEAE Sephadex A-25 equilibrated with 0.1 M Tris-HCl at pH 8.0.
Claim 12
Dependent Claim
37 words
A THF active composition obtainable by extraction from thymus cells by collection of the component eluted at 0.15 M NaCl from an anion exchange chromatograph on DEAE Sephadex A-25 equilibrated with 0.1 M Tris-HCl at pH 8.0.