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Method for the preparation of low molecular weight porcine lympho-reticular polypeptides

a low molecular weight, porcine lympho-reticular technology, applied in the direction of peptidases, peptide sources, enzymology, etc., can solve the problems of unsafe, dangerous, and have the potential to produce severe deleterious effects for doctors and patients

Active Publication Date: 2021-02-18
DE URBINA GAVIRIA SANTIAGO +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes a method for making low molecular weight polypeptides from porcine liver and spleen. The method involves enzymatic hydrolysis using a combination of enzymes with proteolytic and amylase activities. The ratio of liver to spleen can vary depending on the enzymes used. The enzymes are pancreatin, papain, pepsin, or mixtures thereof. The method also includes a cleansing step and the use of a surfactant solution containing benzalkonium chloride. The pH range is maintained through the use of a weak organic acid. The method can be carried out at temperatures of 35° C and 55° C. The technical effect of the patent is to provide a reliable and efficient method for making low molecular weight porcine lympho-reticular polypeptides.

Problems solved by technology

They were dangerous, unsafe and had the potential to produce severe deleterious effects for doctors and patients.

Method used

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  • Method for the preparation of low molecular weight porcine lympho-reticular polypeptides
  • Method for the preparation of low molecular weight porcine lympho-reticular polypeptides
  • Method for the preparation of low molecular weight porcine lympho-reticular polypeptides

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example 1

Preparation of Low Molecular Weight Porcine Lympho-Reticular Polypeptides

[0059]Ingredients / Reagents.

[0060]Fresh porcine liver (certified free of antibiotics and hormones). Fresh porcine spleen (certified free of antibiotics and hormones). Benzalkonium chloride solution (1-2% concentration). Purified water. Papain (diluted in the purified water). Pepsin (anhydrous). Pancreatin. Diastase. Citric acid. Methyl 4-hydroxybenzoate. Propyl 4-hydroxybenzoate.

[0061]Equipment. Stainless steel meat grinder. Stainless steel industrial blender. Stainless steel digester with temperature, pH, and agitation controls. Drying oven with temperature and vacuum controls. Stainless steel drying trays. Stainless steel sieves. Scale. Laboratory glassware. pH meter / controller. Storage bags and containers

[0062]Preparation.

[0063]Approximately 6.6 pounds of fresh porcine livers and 4.4 pounds of fresh porcine spleens, both certified to be free of antibiotics and hormones, are weighed out and trimmed to remove a...

example 2

Evaluation of Cytotoxicity and Anti-Inflammatory Effects of Low Molecular Weight Porcine Lympho-Reticular Polypeptides

[0074]The cytotoxicity and anti-Inflammatory effects of low molecular weight lympho-reticular polypeptides obtained in the Example 1 were evaluated in human cell cultures.

[0075]A Human Acute Monocytic Leukemia cell line—THP-1 was used for the purpose of this study. Cells were maintained in log phase growth in 75 cm2 Falcon cell culture flaks containing complete medium (CM):Gibco RPMI 1640-L-Glutamine medium supplemented with 10% fetal bovine serum and 1000 / mL Gibco Pen Strep. Cell cultures were incubated at 37° C. under 5% CO2 in a humidified atmosphere.

[0076]THP-1 cells were seeded in 96 well plates with 200 μL CM containing 5.0×105 cells and treated with 50 ng / mL PMA for 48 hours to induce differentiation into macrophage-like cells MDM, then washed 4× with CM and allowed to rest for 5 to 7 days with media exchanged for fresh media every two days. MDM were stimulate...

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Abstract

A method for preparation of low molecular weight porcine lympho-reticular polypeptides. The method comprises the enzymatic hydrolysis of a source of protein, wherein the source of protein comprises a blend of porcine liver and porcine spleen, with an enzyme having proteolytic activity and an enzyme having amylase activity.

Description

FIELD OF THE INVENTION[0001]The present invention relates to a method for the preparation of low molecular weight polypeptides. More particularly, the present invention is related to an enzymatic hydrolysis method to produce low molecular weight lympho-reticular polypeptides.BACKGROUND OF THE INVENTION[0002]Since ancient times, proteins of animal and vegetal origin have been used for nutritional and developmental purposes. In fact, diets with normal intake of proteins ensure better development and functioning of physiological functions and organic defenses. Broths with specific protein content, e.g., liver, spleen, bone marrow, and / or blood, have been used for centuries, and as recently as the nineteenth and twentieth centuries, in order to accelerate convalescence in patients affected by a wide variety of pathologies. At the beginning of the twentieth century, various collaborators in Argentina began to use extracts from healthy tissues that they called lysates, obtained from diffe...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12P21/06C07K14/47
CPCC12P21/06C07K14/47C12N9/50C12Y302/01001C12Y301/03008C12Y304/22002C12Y304/23001
Inventor DE URBINA GAVIRIA, SANTIAGODE URBINA, SANTIAGO
Owner DE URBINA GAVIRIA SANTIAGO
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