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Patent US5284777A successfully refreshed from Google Patents.
US5284777A
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Combined glycated hemoglobin and immunoturbidometric glycated albumin assay from whole blood lysate
Assignee: Isolab Inc
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Filing: 1991-03-04
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Granted: 1994-02-08
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Expires: 2011-03-04
Abstract
Abstract A method and kit for the isolation and quantitation of glycated hemoglobin and other glycated proteins, specifically albumin, from a single sample of whole blood. Glycated hemoglobin is calculated from an eluate resulting from the contact of the whole blood hemolysate with a boronated resin. Glycated albumin and other plasma proteins are calculated using immunoturbidimetry, resulting from reaction between a buffered antibody solution and the albumin or other protein in the previous eluate.
Inventors
- 👤 Murray A. Rosenthal
- 👤 David R. Hocking
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 06:41
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/68; G01N33/6803; G01N33/6842; G01N33/72; G01N33/721; G01N33/723; Y; Y10; Y10S; Y10S436/00; Y10S436/825; Y10T; Y10T436/00; Y10T436/10; Y10T436/107497
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
101 words
A method of quantitating a glycated non-hemoglobin protein in a whole blood lysate, said method comprising the steps of: (a) separating glycated proteins from non-glycated proteins by contacting said lysate with a boronated affinity column, to obtain an eluate containing glycated hemoglobin and at least one glycated non-hemoglobin protein; (b) contacting said eluate with an antibody which specifically binds said non-hemoglobin protein, said antibody being in solution buffered to a Ph of from about 7.0 to 9.0, whereby turbidity is produced in said solution; and (c) determining the amount of said non-hemoglobin protein in said turbid solution by measuring said turbidity.
Claim 2
Independent Claim
101 words
A method of quantitating a glycated non-hemoglobin protein in a whole blood lysate, said method comprising the steps of: (a) separating glycated proteins from non-glycated proteins by contacting said lysate with a boronated affinity column, to obtain an eluate containing glycated hemoglobin and at least one glycated non-hemoglobin protein; (b) contacting said eluate with an antibody which specifically binds said non-hemoglobin protein, said antibody being in solution buffered to a Ph of from about 7.0 to 9.0, whereby turbidity is produced in said solution; and (c) determining the amount of said non-hemoglobin protein in said turbid solution by measuring said turbidity.
Claim 3
Dependent Claim
13 words
The method according to claim 1 wherein said glycated non-hemoglobin protein comprises albumin.
Claim 4
Dependent Claim
21 words
The method according to claim 3 wherein said turbidity is determined by measuring the amount of light transmitted through said solution.
Claim 5
Dependent Claim
22 words
The method according to claim 3 wherein said transmitted light is measured at a wavelength of about 305 to about 405 nanometers.
Claim 6
Dependent Claim
22 words
The method according to claim 4 wherein said transmitted light is measured at a wavelength of about 340 to about 380 nanometers.
Claim 7
Dependent Claim
13 words
The method according to claim 1 wherein said antibody is a polyclonal antibody.
Claim 8
Dependent Claim
14 words
The method according to claim 1 wherein said antibody is a goat antihuman antibody.
Claim 9
Dependent Claim
15 words
The method according to claim 1 wherein said antibody specifically binds albumin in said solution.
Claim 10
Independent Claim
154 words
A method of determining amounts of glycated non-hemoglobin protein present in a sample of whole blood containing glycated hemoglobin, said method comprising the steps of: (a) lysing said sample of whole blood; (b) contacting said lysed sample of whole blood with a boronated resin to absorb glycated proteins, said glycated proteins comprising glycated hemoglobin and at least one glycated non-hemoglobin protein; (c) eluting an eluate containing said glycated proteins from said boronated resin; (d) contacting said eluate with an antibody which specifically binds said at least one non-hemoglobin glycated protein, said antibody being in a solution buffered to a pH of from about 7.0 to 9.0, whereby turbidity is produced in said solution; and (e) determining the amount of said at least one glycated non-hemoglobin protein in said solution by measuring said turbidity and comparing said turbidity to that of a standard to quantitate the amount of said at least one glycated non-hemoglobin protein.
Claim 11
Independent Claim
154 words
A method of determining amounts of glycated non-hemoglobin protein present in a sample of whole blood containing glycated hemoglobin, said method comprising the steps of: (a) lysing said sample of whole blood; (b) contacting said lysed sample of whole blood with a boronated resin to absorb glycated proteins, said glycated proteins comprising glycated hemoglobin and at least one glycated non-hemoglobin protein; (c) eluting an eluate containing said glycated proteins from said boronated resin; (d) contacting said eluate with an antibody which specifically binds said at least one non-hemoglobin glycated protein, said antibody being in a solution buffered to a pH of from about 7.0 to 9.0, whereby turbidity is produced in said solution; and (e) determining the amount of said at least one glycated non-hemoglobin protein in said solution by measuring said turbidity and comparing said turbidity to that of a standard to quantitate the amount of said at least one glycated non-hemoglobin protein.
Claim 12
Dependent Claim
26 words
The method according to claim 9 further comprising the step of determining the amount of glycated hemoglobin in said eluate, said step occurring after step (c).
Claim 13
Dependent Claim
16 words
The method according to claim 9 wherein said at least one non-hemoglobin glycated protein is albumin.
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