US4229541A
Expired / Lapsed
US
⏱️ 0.0 years remaining
In vitro cultivation of horseshoe crab amebocytes
Assignee: Baxter International Inc
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Filing: 1978-10-06
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Granted: 1980-10-21
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Expires: 1998-10-06
Abstract
Abstract A culture medium for cultivating horseshoe crab amebocytes, called Limulus amebocytes, composed of inorganic salts, vitamins, amino acids, and carbon sources including glucose is given. The process for cultivating the Limulus amebocytes depends on maintaining a pH between 6.5 and 9.0 while agitating the medium. Production of Limulus lysate from these cells is an eventual goal.
Inventors
- 👤 Frederick C. Pearson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 22:22
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0601; C12Q; C12Q1/00; C12Q1/02; C12Q1/04; C12Q1/045; C12N2500/00; C12N2500/30; C12N2500/32; C12N2500/38
Claims Analysis
Total Claims: 7
Claim 1
Independent Claim
18 words
A culture medium for cultivating Limulus amebocytes, comprising glucose, L-cystine, salts, vitamins, amino acids and horseshoe crab serum.
Claim 2
Dependent Claim
18 words
A culture medium for cultivating Limulus amebocytes, comprising glucose, L-cystine, salts, vitamins, amino acids and horseshoe crab serum.
Claim 3
Dependent Claim
127 words
A culture medium for cultivating Limulus amebocytes in vitro comprising horseshoe crab serum and: NaCl: 1000 mg/L CaCl 2 : 30.0 KCl: 30.0 MgCl 2 .6H 2 O: 2280. MgSO 4 .7H 2 O: 2780. NaHCO 3 : 350 NaH 2 PO 4 .H 2 O: 1013.0 alpha Ketoglutaric acid: 370. Glucose: 1000. Malic acid: 670 L-alanine: 225 L-Arginine HCl: 700.0 L-Asparagine: 350.00 L-Aspartic acid: 350.0 L-Cystine: 100. L-Glutamic acid: 600.00 L-Glutamine: 600.00 Glycine: 650.00 L-Histidine: 2500.00 L-Isoleucine: 50.00 L-Leucine: 75.00 L-Lysine HCl: 625.00 L-Methionine: 50.00 L-Phenylalinine: 150.00 L-Proline: 350.00 DL-Serine: 1100.00 L-Threonine: 175.00 L-Tryptophane: 100.00 L-Tyrosine: 50.00 L-Valine: 100.0 VITAMINS Biotin: 0.01 D-Ca pantothenate: 0.02 Choline chloride: 0.20 Folic acid: 0.02 i-Inositol: 0.02 Niacin: 0.02 Para-aminobenzoic acid: 0.02 Pyridoxine HCl: 0.02 Riboflavin: 0.02 Thiamine HCl: 0.02.
Claim 4
Dependent Claim
127 words
A culture medium for cultivating Limulus amebocytes in vitro comprising horseshoe crab serum and: NaCl: 1000 mg/L CaCl 2 : 30.0 KCl: 30.0 MgCl 2 .6H 2 O: 2280. MgSO 4 .7H 2 O: 2780. NaHCO 3 : 350 NaH 2 PO 4 .H 2 O: 1013.0 alpha Ketoglutaric acid: 370. Glucose: 1000. Malic acid: 670 L-alanine: 225 L-Arginine HCl: 700.0 L-Asparagine: 350.00 L-Aspartic acid: 350.0 L-Cystine: 100. L-Glutamic acid: 600.00 L-Glutamine: 600.00 Glycine: 650.00 L-Histidine: 2500.00 L-Isoleucine: 50.00 L-Leucine: 75.00 L-Lysine HCl: 625.00 L-Methionine: 50.00 L-Phenylalinine: 150.00 L-Proline: 350.00 DL-Serine: 1100.00 L-Threonine: 175.00 L-Tryptophane: 100.00 L-Tyrosine: 50.00 L-Valine: 100.0 VITAMINS Biotin: 0.01 D-Ca pantothenate: 0.02 Choline chloride: 0.20 Folic acid: 0.02 i-Inositol: 0.02 Niacin: 0.02 Para-aminobenzoic acid: 0.02 Pyridoxine HCl: 0.02 Riboflavin: 0.02 Thiamine HCl: 0.02.
Claim 5
Dependent Claim
54 words
A process for propagating Limulus amebocytes in vitro, comprising inoculating Limulus amebocytes into a nutrient medium comprising a carbon source, salts, vitamins, amino acids and horseshoe crab serum, constantly agitating the suspension of amebocytes in the medium at a temperature of from 12° C. to 30° C., and isolating the amebocytes from the suspension.
Claim 6
Dependent Claim
54 words
A process for propagating Limulus amebocytes in vitro, comprising inoculating Limulus amebocytes into a nutrient medium comprising a carbon source, salts, vitamins, amino acids and horseshoe crab serum, constantly agitating the suspension of amebocytes in the medium at a temperature of from 12° C. to 30° C., and isolating the amebocytes from the suspension.
Claim 7
Dependent Claim
129 words
A process of claim 3 wherein the nutrient medium is composed of horseshoe crab serum and: NaCl: 1000 mg/L CaCl 2 : 30.0 KCl: 30.0 MgCl 2 .6H 2 O: 2280. MgSO 4 .7H 2 O: 2780. NaHCO 3 : 350 NaH 2 PO 4 .H 2 O: 1013.0 alpha Ketoglutaric acid: 370. Glucose: 1000. Malic acid: 670 L-Alanine: 225 L-Arginine HCl: 700.0 L-Asparagine: 350.00 L-Aspartic acid: 350.0 L-Cystine: 100. L-Glutamic acid: 600.00 L-Glutamine: 600.00 Glycine: 650.00 L-Histidine: 2500.00 L-Isoleucine: 50.00 L-Leucine: 75.00 L-Lysine HCl: 625.00 L-Methionine: 50.00 L-Phenylalinine: 150.00 L-Proline: 350.00 DL-Serine: 1100.00 L-Threonine: 175.00 L-Tryptophane: 100.00 L-Tyrosine: 50.00 L-Valine: 100.0 VITAMINS Biotin: 0.01 D-Ca pantothenate: 0.02 Choline chloride: 0.20 Folic acid: 0.02 i-Inositol: 0.02 Niacin: 0.02 Para-aminobenzoic acid: 0.02 Pyridoxine HCl: 0.02 Riboflavin: 0.02 Thiamine HCl: 0.02.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1983-10-21 | 1984-04-21 | 1984-10-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-10-21 | 1988-04-21 | 1988-10-21 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-10-21 | 1992-04-21 | 1992-10-21 | $7700.00 | Window Closed / Lapsed |