US4055752A
Expired / Lapsed
US
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Method and apparatus for the determination of enzyme activity
Assignee: Carl Zeiss AG
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Filing: 1975-01-31
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Granted: 1977-10-25
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Expires: 1995-01-31
Abstract
Abstract Enzyme activity for a given sample is automatically determined by measuring light-extinction, at each of a succession of "points" in time and involving a plurality of measurements at each such time. At each of these "points" in time, the several individual measurements are made within a period of time which is brief as compared with the period of time between successive measurement points, and mean value and standard deviation are calculated from these individual measurements. A regression curve is drawn through the measurement points thus obtained, the enzyme activity being determined from the slope thereof at a predetermined time. A total measurement error is also calculated at the same time.
Inventors
- 👤 Walter Kappe
- 👤 Gotz-Reinhard Lampe
- 👤 Harald Neuer
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 21:36
CPC Classifications
G; G01; G01N; G01N21/00; G01N21/17; G01N21/25; G01N21/27; G01N21/272; Y; Y10; Y10S; Y10S435/00; Y10S435/808; Y10T; Y10T436/00; Y10T436/11; Y10T436/115831
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
113 words
The method of determining enzyme activity in a given sample by plural light-extinction measurements on the sample employing electronic digital calculating apparatus which comprises performing a predetermined relatively rapid sequence of plural extinction measurements at a first point in time, repeating said relatively rapid sequence of measurements at second and succeeding relatively greatly spaced points in time, computing via said digital calculating apparatus the mean value to establish a measurement point for the plural measurements of each such rapid sequence, establishing a regression curve through the said measurement points, taking the tangent of said curve at a particular point in time, and calculating from the slope of this tangent the enzyme activity sought.
Claim 2
Independent Claim
113 words
The method of determining enzyme activity in a given sample by plural light-extinction measurements on the sample employing electronic digital calculating apparatus which comprises performing a predetermined relatively rapid sequence of plural extinction measurements at a first point in time, repeating said relatively rapid sequence of measurements at second and succeeding relatively greatly spaced points in time, computing via said digital calculating apparatus the mean value to establish a measurement point for the plural measurements of each such rapid sequence, establishing a regression curve through the said measurement points, taking the tangent of said curve at a particular point in time, and calculating from the slope of this tangent the enzyme activity sought.
Claim 3
Dependent Claim
54 words
The method of claim 1, in which for each measuring point the mean value and the standard deviation are calculated and in which an overall measurement-error is calculated via said digital calculating apparatus from the deviation of said regression curve from the measurement points, said standard deviation and the curvature of the regression curve.
Claim 4
Dependent Claim
41 words
The method as in claim 2 wherein said measurement-error computing step includes the step of determining the square of the difference of the mean value at a measuring point and the value of said established regression curve at said measuring point.
Claim 5
Independent Claim
164 words
The method of determining enzyme activity in a given sample by plural light-extinction measurements on the sample, which comprises performing a predetermined relatively rapid sequence of plural extinction measurements at a first point in time, repeating said relatively rapid sequence of measurements at second and succeeding relatively greatly spaced points in time, computing the mean value to establish a measurement point for the plural measurements of each such rapid sequence, establishing a regression curve through the said measurement points, taking the tangent of said curve at a particular point in time, and calculating from the slope of this tangent the enzyme activity sought in which enzyme activity of each of a plurality of cell-supported samples is measured, by indexing such cells successively into measurement position for the said first-point sequence of measurements on all samples prior to performing the second-point sequence of measurements on all samples, whereby the plural time-spaced sequences of one sample are time-interlaced with the corresponding time-spaced sequences of another sample.
Claim 6
Independent Claim
164 words
The method of determining enzyme activity in a given sample by plural light-extinction measurements on the sample, which comprises performing a predetermined relatively rapid sequence of plural extinction measurements at a first point in time, repeating said relatively rapid sequence of measurements at second and succeeding relatively greatly spaced points in time, computing the mean value to establish a measurement point for the plural measurements of each such rapid sequence, establishing a regression curve through the said measurement points, taking the tangent of said curve at a particular point in time, and calculating from the slope of this tangent the enzyme activity sought in which enzyme activity of each of a plurality of cell-supported samples is measured, by indexing such cells successively into measurement position for the said first-point sequence of measurements on all samples prior to performing the second-point sequence of measurements on all samples, whereby the plural time-spaced sequences of one sample are time-interlaced with the corresponding time-spaced sequences of another sample.
Claim 7
Independent Claim
234 words
The method of determining enzyme activity in a given sample by plural light-extinction measurements on the sample, which comprises performing a predetermined relatively rapid sequence of plural extinction measurements at a first point in time, repeating said relatively rapid sequence of measurements at second and succeeding relatively greatly spaced points in time, computing the mean value to establish a measurement point for the plural measurements of each such rapid sequence, establishing a regression curve through the said measurement points, taking the tangent of said curve at a particular point in time and calculating from the slope of this tangent the enzyme activity sought in which enzyme activity of each of a plurality of cell-supported samples is measured, by indexing such cells successively into measurement position for the said first-point sequence of measurements on all samples prior to performing the second-point sequence of measurements on all samples, whereby the plural time-spaced sequences of one sample are time-interlaced with the corresponding time-spaced sequences of another sample, in which the light-extinction measurements include photometrically and adjustably variable amplification of the electric signal, generating and separately storing an optimum amplification level uniquely applicable to each sample observed for the first point-in-time measurement applicable thereto, and synchronously commutating the respective stored levels to correspondingly adjust the amplification level used for optimal measurement of each sample for each of the succeeding points of time at which its light-extinction is measured.
Claim 8
Independent Claim
234 words
The method of determining enzyme activity in a given sample by plural light-extinction measurements on the sample, which comprises performing a predetermined relatively rapid sequence of plural extinction measurements at a first point in time, repeating said relatively rapid sequence of measurements at second and succeeding relatively greatly spaced points in time, computing the mean value to establish a measurement point for the plural measurements of each such rapid sequence, establishing a regression curve through the said measurement points, taking the tangent of said curve at a particular point in time and calculating from the slope of this tangent the enzyme activity sought in which enzyme activity of each of a plurality of cell-supported samples is measured, by indexing such cells successively into measurement position for the said first-point sequence of measurements on all samples prior to performing the second-point sequence of measurements on all samples, whereby the plural time-spaced sequences of one sample are time-interlaced with the corresponding time-spaced sequences of another sample, in which the light-extinction measurements include photometrically and adjustably variable amplification of the electric signal, generating and separately storing an optimum amplification level uniquely applicable to each sample observed for the first point-in-time measurement applicable thereto, and synchronously commutating the respective stored levels to correspondingly adjust the amplification level used for optimal measurement of each sample for each of the succeeding points of time at which its light-extinction is measured.
Claim 9
Independent Claim
120 words
The method of determining enzyme activity in a given sample by plural light-extinction measurements on the sample, which comprises performing a predetermined relatively rapid sequence of plural extinction measurements at a first point in time, repeating said relatively rapid sequence of measurements at second and succeeding relatively greatly spaced points in time, computing the mean value to establish a measurement point for the plural measurements of each such rapid sequence, establishing a regression curve through the said measurement points, taking the tangent of said curve at a particular point in time, and calculating from the slope of this tangent the enzyme activity sought, in which clock-pulse signals are used to determine precisely-spaced like periods of time between said measurement points.
Claim 10
Independent Claim
120 words
The method of determining enzyme activity in a given sample by plural light-extinction measurements on the sample, which comprises performing a predetermined relatively rapid sequence of plural extinction measurements at a first point in time, repeating said relatively rapid sequence of measurements at second and succeeding relatively greatly spaced points in time, computing the mean value to establish a measurement point for the plural measurements of each such rapid sequence, establishing a regression curve through the said measurement points, taking the tangent of said curve at a particular point in time, and calculating from the slope of this tangent the enzyme activity sought, in which clock-pulse signals are used to determine precisely-spaced like periods of time between said measurement points.
Claim 11
Independent Claim
114 words
The method of determining enzyme activity in a given sample by plural light-extinction measurements on the sample, which comprises performing a predetermined relatively rapid sequence of plural extinction measurements at a first point in time, repeating said relatively rapid sequence of measurements at second and succeeding relatively greatly spaced points in time, computing the mean value to establish a measurement point for the plural measurements of each such rapid sequence, establishing a regression curve through the said measurement points, taking the tangent of said curve at a particular point in time, and calculating from the slope of this tangent the enzyme activity sought, in which the regression curve is selected as a second-degree polynomial.
Claim 12
Independent Claim
114 words
The method of determining enzyme activity in a given sample by plural light-extinction measurements on the sample, which comprises performing a predetermined relatively rapid sequence of plural extinction measurements at a first point in time, repeating said relatively rapid sequence of measurements at second and succeeding relatively greatly spaced points in time, computing the mean value to establish a measurement point for the plural measurements of each such rapid sequence, establishing a regression curve through the said measurement points, taking the tangent of said curve at a particular point in time, and calculating from the slope of this tangent the enzyme activity sought, in which the regression curve is selected as a second-degree polynomial.
Claim 13
Dependent Claim
111 words
Apparatus for light-extinction measurement of enzyme activity in a sample, comprising a light source, photometer means spaced from and oriented for electrical response to light from said source, sample-supporting means interposed between said source and photometer means, analog-digital converter means connected to the electrical output of said photometer means, computer means connected to control a predetermined rapid sequence of measurements at each of a plurality of spaced points in time, said converter means being connected to said computer means, and said computer means including means for computing and storing the mean value for the collective measurements of each such sequence and for computing the enzyme activity using said stored measurement values.
Claim 14
Dependent Claim
111 words
Apparatus for light-extinction measurement of enzyme activity in a sample, comprising a light source, photometer means spaced from and oriented for electrical response to light from said source, sample-supporting means interposed between said source and photometer means, analog-digital converter means connected to the electrical output of said photometer means, computer means connected to control a predetermined rapid sequence of measurements at each of a plurality of spaced points in time, said converter means being connected to said computer means, and said computer means including means for computing and storing the mean value for the collective measurements of each such sequence and for computing the enzyme activity using said stored measurement values.
Claim 15
Dependent Claim
22 words
Apparatus according to claim 8, in which said computer means are connected to a real-time-clock for timing the sequence of said measurements.
Claim 16
Dependent Claim
77 words
Apparatus according to claim 8, in which said photometer means includes variable-gain amplifier means, said computer including means for calculating from measured output for measurements at a first point in time an optimum gain setting for said amplifier means, means for storing a value corresponding to said optimum setting, and means establishing a control connection from said storage means to reset said amplifier gain for measurements at a later point in time based on the stored value.
Claim 17
Dependent Claim
24 words
Apparatus according to claim 10, in which the variable gain of said amplifier means comprises a plurality of discrete, purely digitally selectable amplification stages.
Claim 18
Dependent Claim
24 words
Apparatus according to claim 8, characterized by the fact that said photometer means includes coded filters whose code numbers are interrogated by the computer.
Claim 19
Dependent Claim
34 words
Apparatus as in claim 8, wherein said computer means includes means for determining the standard deviation at said spaced points in time, and means for fitting a curve through said mean and deviation measurements.
Claim 20
Dependent Claim
26 words
Apparatus as in claim 13 wherein said computer means further comprises means for determining the derivative of said curve produced by said curve fitting portion thereof.
Claim 21
Dependent Claim
108 words
In combination in enzyme activity determining means, light source and light detector means for determining relative light transmissivity of a sample disposed therebetween, means for repetitively sampling the output of said detector means at spaced points in time relatively large in comparison to the inter-sample interval for any said point in time, computer means for receiving said sampled detector output signals, said computer means including means for determining statistical light transmissivity for the samples at any point in time, means for fitting a curve to said statistical properties determined by said means therefor, and means for determining the derivative of said curve determined by said curve fitting means.
Claim 22
Dependent Claim
108 words
In combination in enzyme activity determining means, light source and light detector means for determining relative light transmissivity of a sample disposed therebetween, means for repetitively sampling the output of said detector means at spaced points in time relatively large in comparison to the inter-sample interval for any said point in time, computer means for receiving said sampled detector output signals, said computer means including means for determining statistical light transmissivity for the samples at any point in time, means for fitting a curve to said statistical properties determined by said means therefor, and means for determining the derivative of said curve determined by said curve fitting means.
Claim 23
Dependent Claim
17 words
A combination as in claim 15 wherein said statistical property determining means includes mean value determining means.
Claim 24
Dependent Claim
24 words
A combination as in claim 16 wherein said curve fitting means includes means for fitting a second degree polynomial curve to said mean values.
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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) | 1980-10-24 | 1981-04-25 | 1981-10-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1984-10-24 | 1985-04-25 | 1985-10-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1988-10-24 | 1989-04-25 | 1989-10-25 | $7700.00 | Window Closed / Lapsed |