Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for separating unmodified hemoglobin from cross-linked hemoglobin

a technology of unmodified hemoglobin and cross-linked hemoglobin, which is applied in the field of separating unmodified hemoglobin from cross-linked hemoglobin, can solve the problems of patient death, complex patient recovery, and a significant fraction of unmodified tetrameric hemoglobin, and achieves the reduction or elimination of significant renal elimination of hemoglobin, improve intravascular retention time, and suitable oncotic pressure

Inactive Publication Date: 2005-06-30
OPK BIOTECH
View PDF24 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0009] The advantages of this invention include providing a blood-substitute with an improved intravascular retention time, a reduction or elimination of significant renal elimination of hemoglobin and the side effects associated therewith, a suitable oncotic pressure, and reduced hypertensive effects.

Problems solved by technology

For example, the use of whole blood often is accompanied by the risk of transmission of hepatitis-producing viruses and AIDS-producing viruses which can complicate patient recovery or result in patient fatalities.
However, solutions of cross-linked hemoglobin still typically contain a significant fraction of unmodified tetrameric hemoglobin.
Furthermore, current means for separation, such as standard filtration, do not adequately distinguish between unmodified tetrameric hemoglobin and modified tetrameric hemoglobin.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for separating unmodified hemoglobin from cross-linked hemoglobin

Examples

Experimental program
Comparison scheme
Effect test

example i

Diafiltration of Deoxygenated Hb Solution Containing a Higher Concentration Dissociation Buffer

[0069] A polymerized Hb solution was formed according to the method described in Example I of U.S. Pat. No. 5,084,558, issued to Rausch et al. This Hb solution was analyzed by gel permeation chromatography (GPC) and found to comprise about 45% Hb dimers, about 15% unmodified Hb tetramers, and about 40% polymerized Hb molecules which were larger than unmodified tetramers. One liter of a dissociation buffer containing 1.5 M MgCl2, 0.1 M Bis-Tris and 0.2 mM EDTA (pH 6.5) was added to one liter of the Hb solution. This mixture was then recirculated through a 100 kD polysulfone ultrafilter (Millipore Catalog No. PTHK 000C5) to concentrate the mixture to a volume of one liter. The concentrated mixture was subsequently diafiltered with 11 volumes of a dissociation buffer comprising 0.7 M MgCl2, 0.05 M Bis-Tris and 0.1 mM EDTA (pH 6.5). The filtered Hb solution was then washed and equilibrated wi...

example ii

Diafiltration of Deoxygenated Hb Solution Containing a Lower Concentration Dissociation Buffer

[0071] One hundred seventy milliliters (ml) of a dissociation buffer containing 0.75 M MgCl2, 0.05 M Bis-Tris and 0.1 mM EDTA (pH 7.5) was added to 15 ml of the initial polymerized Hb solution of Example I and then 15 ml of a two-fold concentrate of the dissociation buffer was added. The Hb solution was then recirculated through a Chemineer, Inc. / Kenics static mixer and then diafiltered by a 100 kD ultrafilter (Amicon YM 100, Catalog No. 14451) to obtain 200 ml of Hb solution.

[0072] The Hb solution was then diafiltered with 3 volume exchanges of the dissociation buffer and lastly washed and equilibrated with a deoxygenated buffer containing 27 mM sodium lactate, 12 mM N-acetyl cysteine, 115 mM NaCl, 4 mM KCl, and 1.36 mM CaCl2 in WFI. The molecular weight distribution of the resulting Hb solution was then analyzed by GPC.

[0073] The results of these analyses are shown in FIG. 2. The Hb so...

example iii

Diafiltration of Oxygenated and Deoxygenated Hb Solutions Without a Dissociation Buffer

[0074] A polymerized Hb solution was formed according to the method described in Example 1 of U.S. Pat. No. 5,955,581, issued to Rausch et al. This Hb solution was analyzed by GPC and found to comprise about 3.5% Hb dimers, 31% unmodified Hb tetramers and about 65.5% polymerized Hb molecules which were larger than unmodified tetramers.

[0075] Two liters of the Hb solution were oxygenated through an oxygenation cartridge with a gaseous mixture, comprising 98% oxygen and 2% carbon dioxide, until 95% oxygenated Hb valves were obtained by a co-oximeter (Co-Oximeter Model #482; Instrumentation Laboratory, Lexington, Mass.).

[0076] The oxygenated Hb solution was then diafiltered with 7 volumes of an oxygenated buffer solution containing 27 mM solution lactate, 12 mM N-acetyl-L-cysteine, 115 mM NaCl, 4 mM KCl and 1.4 mM CaCl2 in WFI against a 100 kD ultrafilter. Throughout this process, the Hb solution ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
temperatureaaaaaaaaaa
temperatureaaaaaaaaaa
temperatureaaaaaaaaaa
Login to View More

Abstract

A method for separating unmodified hemoglobin from cross-linked hemoglobin in a hemoglobin solution. The method involves contacting the hemoglobin solution with a least one dissociating agent to form a dissociation solution wherein unmodified tetrameric hemoglobin is dissociated to form hemoglobin dimers. The hemoglobin dimers are then separated from the dissociation solution, while retaining the cross-linked hemoglobin in the dissociation solution.

Description

RELATED APPLICATIONS [0001] This application is a Continuation of U.S. Ser. No. 10 / 420,630 filed on Apr. 18, 2003, which is a Continuation of U.S. Ser. No. 09 / 460,153, filed on Dec. 13, 1999, which is a Continuation of U.S. Ser. No. 09 / 305,412, filed on May 5, 1999, which is a Continuation of U.S. Ser. No. 08 / 477,916, filed on Jun. 7, 1995. The entire teachings of the above applications are incorporated herein by reference.BACKGROUND OF THE INVENTION [0002] There exists a need for a blood-substitute to treat or prevent hypoxia resulting from blood loss (e.g, from acute hemorrhage or during surgical operations), resulting from anemia (e.g., pernicious anemia or sickle cell anemia), or resulting from shock (e.g, volume deficiency shock, anaphylactic shock, septic shock or allergic shock). [0003] The use of blood and blood fractions as in these capacities as a blood-substitute is fraught with disadvantages. For example, the use of whole blood often is accompanied by the risk of transmi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(United States)
IPC IPC(8): C07K14/805C07K1/14
CPCC07K14/805
Inventor LIGHT, WILLIAM R.GAWRYL, MARIA S.LACCETTI, ANTHONY J.HOUTCHENS, ROBERT A.
Owner OPK BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com