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US3915648A Expired / Lapsed US ⏱️ 0.0 years remaining

Fluorescence protein and peptide analyzer

Assignee: F Hoffmann La Roche AG
•
Filing: 1973-10-24
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Granted: 1975-10-28
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Expires: 1993-10-24
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Abstract

Abstract A fluorescence-based analyzer, which is readily adaptable to protein or peptide analysis. The adaptability of the analyzer is derived from a novel rotating sample loopholder. In the protein analysis mode, this sample loopholder is provided with a number of loops made of a semi-permeable membrane and the sample loop holder is immersed within a dialysis solution. Thus, a protein containing sample is dialyzed to remove low-molecular weight contaminants which can interfere with the fluorescence measurement. When the instrument is in the peptide analysis mode the aforesaid sample loops are made out of inert, non-porous plastic capable of withstanding strong alkali and elevated temperatures, i.e., 100*C., and the peptide or protein samples contained therein are hydrolyzed to the constituent amino acids.

Inventors

  • 👤 Stanley Stein

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-02 09:14
CPC Classifications
G; G01; G01N; G01N35/00; B; B01; B01D; B01D61/00; B01D61/24; B01D61/28; B01D63/00; B01D63/06; B01D71/00; B01D71/06; B01D71/08; B01D71/10

Claims Analysis

Total Claims: 10
Claim 1
Independent Claim 319 words
AN ANALYZER SYSTEM WHICH IS ADAPTABLE TO THE FLUORESCENCE BASED ASSAY OF EITHER PROTEIN OR PEPTIDE CONTAINING SAMPLES WHICH SYSTEM COMPRISES IN COMBINATION: A. SAMPLE STREAM PROVIDING MEANS WHICH SAMPLE STREAM CONTAINS EITHER PROTEIN OR PEPTIDE COMPOUNDS TO BE ASSAYED, B. SAMPLE LOOPHOLDER MEANS ADAPTABLE TO BE IN A DIALYSIS OR HYDROLYSIS MODE, SAID SAMPLE LOOPHOLDER MEANS COMPRISING: I. AN OUTER ENCLOSURE DEFINING AN INTERNAL CHAMBER WHICH CHAMBER CONTAINS DIALYSIS SOLUTION IN SAID DIALYSIS MODE AND HEATING MEANS IN SAID HYDROLYSIS MODE, AND II. A SAMPLE LOOPHOLDER COMPONENT COMPRISING A STEPPING MOTOR MEANS, SHAFT MEANS CONNECTED TO SAID STEPPING MOTOR MEANS, A STATOR ELEMENT HAVING TWO RADIALLY DISPOSED CHANNEL PAIRS AND A ROTATOR ELEMENT CONNECTED TO SAID SHAFT MEANS HAVING A PLURALITY OF CHANNEL PAIRS RADIALLY AND AND SYMMETRICALLY DISPOSED ABOUT ITS CENTRAL AXIS, SAID PLURALITY OF CHANNEL PAIRS BEING SO ARRANGED AND CONSTRUCTED AS TO SERIALLY INDEX WITH EACH OF SAID CHANNEL PAIRS IN SAID STATOR ELEMENTS AS SAID ROTATOR IS ROTATED RELATIVE TO SAID STATOR, EACH OF SAID ROTATOR CHANNEL PAIRS FURTHER HAVING A SAMPLE LOOP BRINDING THE TWO CHANNELS IN EACH PAIR, SAID SAMPLE LOOP BEING SELECTED FROM THE GROUP CONSISTING OF A SEMIPERMEABLE CELLULOSE TUBE FOR THE DIALYSIS MODE AND AN INERT, NON-POROUS, HEAT AND ALKALI STABLE PLASTIC FOR THE HYDROLYSIS MODE, SAID ROTATOR AND SAID STATOR BEING HELD TOGETHER TIGHTLY TO FORM A LIQUID TIGHT SEAL BETWEEN THEM WITH SAID ROTATOR BEING ROTATABLE WITH RESPECT TO SAID STATOR, AND C. FLUORESCENCE DETECTION MEANS, WHEREIN SAID SAMPLE STREAM PROVIDING MEANS IS OPERATIVELY CONNECTED TO ONE CHANNEL OF ONE OF SAID CHANNEL PAIRS OF SAID STATOR SO AS TO PROVIDE A SAMPLE STREAM TO SAID CHANNEL AND ANY CHANNEL OF SAID ROTATOR WHICH IS INDEXED THEREWITH AND SAID FLUORESCENCE DETECTION MEANS IS OPERATIVELY CONNECTED TO ONE CHANNEL OF THE OTHER OF SAID CHANNEL PAIRS SO AS TO OBTAIN A DIALYZED OR HYDROLYZED SAMPLE STREAM FOR ASSAY.
Claim 2
Dependent Claim 319 words
AN ANALYZER SYSTEM WHICH IS ADAPTABLE TO THE FLUORESCENCE BASED ASSAY OF EITHER PROTEIN OR PEPTIDE CONTAINING SAMPLES WHICH SYSTEM COMPRISES IN COMBINATION: A. SAMPLE STREAM PROVIDING MEANS WHICH SAMPLE STREAM CONTAINS EITHER PROTEIN OR PEPTIDE COMPOUNDS TO BE ASSAYED, B. SAMPLE LOOPHOLDER MEANS ADAPTABLE TO BE IN A DIALYSIS OR HYDROLYSIS MODE, SAID SAMPLE LOOPHOLDER MEANS COMPRISING: I. AN OUTER ENCLOSURE DEFINING AN INTERNAL CHAMBER WHICH CHAMBER CONTAINS DIALYSIS SOLUTION IN SAID DIALYSIS MODE AND HEATING MEANS IN SAID HYDROLYSIS MODE, AND II. A SAMPLE LOOPHOLDER COMPONENT COMPRISING A STEPPING MOTOR MEANS, SHAFT MEANS CONNECTED TO SAID STEPPING MOTOR MEANS, A STATOR ELEMENT HAVING TWO RADIALLY DISPOSED CHANNEL PAIRS AND A ROTATOR ELEMENT CONNECTED TO SAID SHAFT MEANS HAVING A PLURALITY OF CHANNEL PAIRS RADIALLY AND AND SYMMETRICALLY DISPOSED ABOUT ITS CENTRAL AXIS, SAID PLURALITY OF CHANNEL PAIRS BEING SO ARRANGED AND CONSTRUCTED AS TO SERIALLY INDEX WITH EACH OF SAID CHANNEL PAIRS IN SAID STATOR ELEMENTS AS SAID ROTATOR IS ROTATED RELATIVE TO SAID STATOR, EACH OF SAID ROTATOR CHANNEL PAIRS FURTHER HAVING A SAMPLE LOOP BRINDING THE TWO CHANNELS IN EACH PAIR, SAID SAMPLE LOOP BEING SELECTED FROM THE GROUP CONSISTING OF A SEMIPERMEABLE CELLULOSE TUBE FOR THE DIALYSIS MODE AND AN INERT, NON-POROUS, HEAT AND ALKALI STABLE PLASTIC FOR THE HYDROLYSIS MODE, SAID ROTATOR AND SAID STATOR BEING HELD TOGETHER TIGHTLY TO FORM A LIQUID TIGHT SEAL BETWEEN THEM WITH SAID ROTATOR BEING ROTATABLE WITH RESPECT TO SAID STATOR, AND C. FLUORESCENCE DETECTION MEANS, WHEREIN SAID SAMPLE STREAM PROVIDING MEANS IS OPERATIVELY CONNECTED TO ONE CHANNEL OF ONE OF SAID CHANNEL PAIRS OF SAID STATOR SO AS TO PROVIDE A SAMPLE STREAM TO SAID CHANNEL AND ANY CHANNEL OF SAID ROTATOR WHICH IS INDEXED THEREWITH AND SAID FLUORESCENCE DETECTION MEANS IS OPERATIVELY CONNECTED TO ONE CHANNEL OF THE OTHER OF SAID CHANNEL PAIRS SO AS TO OBTAIN A DIALYZED OR HYDROLYZED SAMPLE STREAM FOR ASSAY.
Claim 3
Dependent Claim 32 words
The analyzer system of claim 1 wherein said surfaces between said rotator and said stator are machined to a low friction finish so as to provide said liquid tight seal between them.
Claim 4
Dependent Claim 23 words
The analyzer system of claim 1 wherein said stepping motor has a step corresponding to the number of channel pairs in said rotator.
Claim 5
Dependent Claim 21 words
The analyzer system of claim 1 wherein said sample loops are detachably attached to said channels in a liquid tight fit.
Claim 6
Dependent Claim 159 words
The analyzer system of claim 1 wherein in said hydrolysis mode said sample stream providing means comprises chromatographic column means, first valve means having a collecting and a sampling position, fraction collector means, and alkaline addition means comprising alkaline solution reservoir means, pump means and second valve means having an addition and a recycle position; wherein a sample stream from said chromatographic column means is provided said fraction collector means when said first valve means is in said collecting position and is provided to said sample loopholder means when said first valve means is in said sampling position, wherein further said second valve means is in recycle position when said first valve means is in collecting position and said second valve means is in addition position when said first valve means is in sampling position so that a desired amount of alkaline solution from said reservoir means is added to said sample stream being provided to said sample loopholder.
Claim 7
Dependent Claim 73 words
The analyzer system of claim 1 wherein in said dialysis mode said sample stream providing means comprises chromatographic column means, valve means having a collecting and a sampling position, and fraction collector means, wherein a sample stream from said chromatographic column means is provided said fraction collector means when said valve means is in collecting position and is provided to said sample loopholder means when said valve means is in its sampling position.
Claim 8
Dependent Claim 81 words
The analyzer of claim 1 wherein said fluorescence detection means comprises fluorogen solution reservoir means, pump means, fluorometer means and recorder means whereby the hydrolyzed or dialyzed sample stream from said sample loopholder means is treated with said fluorogen solution so as to form a fuorophor in said sample stream which is excited and the resulting fluorescence measured in said fluorometer means and a signal proportional to said measured fluorescence is given to said recorder means where it is graphically displayed.
Claim 9
Dependent Claim 29 words
The analyzer system of claim 1 wherein in said dialysis mode said sample stream providing means comprises sample injection means for introducing samples manually into said sample loopholder means.
Claim 10
Dependent Claim 14 words
The analyzer system of claim 8 wherein said sample injection means comprises a syringe.

References Cited (Backward Citations)

US2846121A Cited
US3182043A Cited
US3222135A Cited
US3567389A Cited
US3591667A Cited
US3763374A Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1978-10-27 1979-04-28 1979-10-28 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1982-10-27 1983-04-28 1983-10-28 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1986-10-27 1987-04-28 1987-10-28 $7700.00 Window Closed / Lapsed

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