EP0156588A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Automated polypeptide synthesis apparatus
Assignee: Applied Biosystems Inc
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Filing: 1985-03-13
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Granted: —
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Expires: 2005-03-13
Abstract
Abstract An apparatus is provided for automatically constructing a polypeptide of high purity, up to 50 amino acids in length, using only single couplings. The apparatus includes an ac- ivation system for receiving protected amino acids, one kind at a time, having a common vessel (an activator vessel) in which to activate each of the amino acids. Also included is ; reaction vessel for containing a resin used in solid-phase I eptide synthesis for attaching a peptide chain thereto. A transfer system is also provided, which operates under con- rol of a computer, to transfer the activated species from the ctivation system to the reaction vessel and to transfer amino acids, reagents, gases, and solvents from one part of the apparatus to another. The activator system also includes a temperature controlled concentrator vessel in which an activator solvent is replaced by a coupling solvent to enhance the coupling of the activated species to the peptide chain in the reaction vessel. Also included in the synthesizer system is a vortexer for affecting total washing of materials in the .reaction vessel and the reaction vessel itself, an automated peptide resin sampling system, and an autodelivery system for providing individual containers of amino acid to the synthesizer in the order desired in the peptide seauence. A liquid sensor system is also included to monitor transitions between gases and liquids in specific tubes in the synthesizer in order to provide input signals to the computer system for control purposes. The computer system software which controls the operation of the synthesizer is organized according to a series of menus which allows the user of the system to select individual cycles of operation for each vessel in the synthesizer. In addition, an algorithm has been developed which provides for optimum efficiency in the production of a peptide for any given selection of cycles.
Inventors
- 👤 John Bridgham
- 👤 Timothy G. Geiser
- 👤 Michael W. Hunkapiller
- 👤 Stephen B.H. Kent
- 👤 Mark P. Marriott
- 👤 Paul Oliver Ramstad
- 👤 Eric Scott Nordman
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-07 20:54
CPC Classifications
C; C07; C07K; C07K1/00; C07K1/04; C07K1/045; B; B01; B01J; B01J19/00; B01J19/0046; B01J2219/00; B01J2219/00274; B01J2219/00277; B01J2219/00279; B01J2219/00281; B01J2219/00286; B01J2219/00351; B01J2219/00389; B01J2219/00479; B01J2219/00484; B01J2219/00488; B01J2219/0049; B01J2219/00495; B01J2219/00497; B01J2219/005; B01J2219/00583; B01J2219/0059; B01J2219/00596; B01J2219/0068; B01J2219/00686; B01J2219/00689; B01J2219/00695; B01J2219/00718; B01J2219/0072; B01J2219/00725; C40; C40B; C40B40/00; C40B40/04; C40B40/10; C40B60/00; C40B60/14
Claims Analysis
Total Claims: 32
Claim 1
Independent Claim
133 words
A process for the manufacture of peptides by solid phase synthesis in which successive amino acids are brought to a carboxyl-activated state and linked successively together at active sites on a solid substrate via an end carboxyl group, the amino acids being added with the amino group protected and subsequently deprotected after linkage characterised in that: a) the linkage reaction takes place successively in a common reaction vessel (15) containing the substrate b) the reaction vessel is fed successively with an aliquot of the appropriate amino acid in a carboxyl-activated state in an appropriate coupling medium c) the appropriate aliquot of each amino acid in its activated state in said coupling medium is prepared immediately prior to use d) the cycle of process steps together with ancilliary preparatory steps is carried out automatically.
Claim 2
Dependent Claim
133 words
A process for the manufacture of peptides by solid phase synthesis in which successive amino acids are brought to a carboxyl-activated state and linked successively together at active sites on a solid substrate via an end carboxyl group, the amino acids being added with the amino group protected and subsequently deprotected after linkage characterised in that: a) the linkage reaction takes place successively in a common reaction vessel (15) containing the substrate b) the reaction vessel is fed successively with an aliquot of the appropriate amino acid in a carboxyl-activated state in an appropriate coupling medium c) the appropriate aliquot of each amino acid in its activated state in said coupling medium is prepared immediately prior to use d) the cycle of process steps together with ancilliary preparatory steps is carried out automatically.
Claim 3
Dependent Claim
22 words
A process according to claim 1 wherein activation of the amino acids involves production of the corresponding alpha amino protected symmetric anhydride.
Claim 4
Dependent Claim
40 words
A process according to claim 1 or claim 2 which is controlled by a computer programme in which the steps are carried out according to a cycle in which temperatures and cycle periods can be varied with each amino acid.
Claim 5
Dependent Claim
23 words
A process according to any preceding claim in which amino acid reagents are indexed in the series required and monitored by bar code.
Claim 6
Dependent Claim
20 words
A process according to any preceding claim in which the final peptide is separated from the substrate by vortex action.
Claim 7
Dependent Claim
39 words
A process according to any preceding claim in which the activated amino acid is transferred to a concentrator vessel (13) for concentration and replacement of solvent by the coupling medium before transfer to a reaction vessel containing said substrate.
Claim 8
Dependent Claim
97 words
Apparatus for carrying out the process of claim 1 comprising a reaction vessel (15) for accommodating the substrate in an appropriate solvent activation means including an activation vessel (11) and solvent coupling medium substitution means (33, 41) transfer means (Fig. 1B) arranged for successively feeding the reaction vessel with deprotection reagent and activated amino acids in a controlled sequence and for feeding the activation vessel with the appropriate amino acid reagents, solvents and rinsing fluids in controlled sequence under automated control, and computer means (Fig. 2) enabling control of said transfer means according to a programmed cycle.
Claim 9
Dependent Claim
20 words
Apparatus according to claim 7 including computer software enabling temperatures and cycle periods to be varied with each amino acid.
Claim 10
Dependent Claim
35 words
Apparatus according to claim 7 wherein said reaction vessel (15) includes agitation means (48) operating with a vortex action for preventing agglomeration of resin during reaction with each of said activated species of amino acid.
Claim 11
Dependent Claim
18 words
Apparatus according to claim 9 wherein said reaction vessel comprises a container having an axis of rotational symmetry.
Claim 12
Dependent Claim
39 words
Apparatus according to claim 10 wherein said agitation means comprises rotation means arranged for causing one end of said container to execute a circular motion about said axis of rotational symmetry while the other end is held generally stationary.
Claim 13
Dependent Claim
62 words
Apparatus according to any of claims 1 to 10 wherein said activation means includes a concentrator vessel (13) for receiving said activator solvent and each of said activated species of amino acid, one amino acid at a time, from said activator vessel, and said transfer means is arranged for replacing said activator solvent medium from said concentrator vessel by a coupling medium.
Claim 14
Dependent Claim
82 words
Apparatus according to any of claims 7 to 12 wherein said transfer means also includes autodelivery means (Fig. 3) under control of said computer for supplying said amino acids, one kind at a time, to said activation means, in the order desired in said peptide, said autodelivery means including means for indexing individual containers of amino acids in the order desired in said peptide, and extraction means (121,. 123) for extracting the contents of said individual containers, one container at a time.
Claim 15
Dependent Claim
34 words
Apparatus according to claim 13 wherein said extraction means includes needle means (121, 123) for supplying solvent into each of said containers and for extracting said solvent together with amino acid from said containers.
Claim 16
Dependent Claim
31 words
Apparatus according to claim 13 or 14 including individual containers (105) held in said indexing means, each of said containers having a bottom which internally has the shape of a trough.
Claim 17
Dependent Claim
19 words
Apparatus according to claim 15 wherein externally the said containers (145) are adapted to stand upright without external constraints.
Claim 18
Dependent Claim
65 words
Apparatus according to claim 12 wherein said transfer means includes a first tube between said activator vessel and said concentrator vessel and a first valve block means (23) controlled by said computer for controlling flow from said activator vessel to said first tube, and a second valve block means (33) controlled by said computer for controlling flow into said concentrator vessel from said first tube.
Claim 19
Dependent Claim
58 words
Apparatus according to claim 17 wherein said transfer means also includes a second tube between said concentrator vessel and said reaction vessel and a third valve block means (45) controlled by said computer for controlling flow into said reaction vessel, and wherein said second valve block means also controls flow from said concentrator vessel into said second tube.
Claim 20
Dependent Claim
26 words
Apparatus according to claim 17 or 18 including liquid sensor means (27, 40, 48) for detecting transitions between gases and liquids flowing in said first tube.
Claim 21
Dependent Claim
23 words
Apparatus according to claims- 7 to 19 including resin sampler means (59) controlled by said computer for extracting materials from said reaction vessel.
Claim 22
Dependent Claim
64 words
Apparatus according to any of claims 7 to 20 wherein said computer means includes menu means categorizing physical and chemical operations in said device, for permitting control over said operations by an operator, said categorization being segregated into a set of cycles, each cycle being characteristic of a complete set operations in one of said activator vessel, said concentrator vessel, and said reaction vessel.
Claim 23
Dependent Claim
56 words
Apparatus according to claim 21 wherein said computer means includes memory means for storing an algorithm for calculating operating times of said first, second and third valve block means to maximize the production rate of a polypeptide given any particular sequence of cycles in each of said activator vessel, said concentrator vessel, and said reaction vessel.
Claim 24
Dependent Claim
35 words
Apparatus according to claim 22 wherein said computer means includes means for calculating said operating times and switch means for operating said first, second, and third valve block means in accordance with said operating times.
Claim 25
Dependent Claim
114 words
A resin sampler (59) for sampling a reaction vessel of a solid state peptide synthesis comprising: a reservoir; a membrane located in said reservoir for stopping flow of materials above a certain size past the location of said membrane in said reservoir; a first connecting tube connected to said reservoir on one side of said membrane; a first valve for controlling the flow through said connecting tube; a second connecting tube connected to said reservoir on the same side of said membrane as said first connecting tube; a second valve for controlling the flow through said second connecting tube; a third connecting tube connected to said reservoir on the opposite side of said membrane.
Claim 26
Dependent Claim
114 words
A resin sampler (59) for sampling a reaction vessel of a solid state peptide synthesis comprising: a reservoir; a membrane located in said reservoir for stopping flow of materials above a certain size past the location of said membrane in said reservoir; a first connecting tube connected to said reservoir on one side of said membrane; a first valve for controlling the flow through said connecting tube; a second connecting tube connected to said reservoir on the same side of said membrane as said first connecting tube; a second valve for controlling the flow through said second connecting tube; a third connecting tube connected to said reservoir on the opposite side of said membrane.
Claim 27
Dependent Claim
139 words
A liquid sensor for detecting translations between gases and liquids in a translucent tube comprising: two substantially flat substrates (217, 218) constructed of a resilient material held in a substantially opposed position relative to each other and a fixed distance apart near a first end; holding means (229) located opposite said first end and between said substrates for holding said tube, the thickness of said tube and said holding means together exceeding said fixed distance; a light means (270) held by said holder means for providing radiant energy from one side of said tube; detector means (271) held by said holder means on the opposite side of said tube from said light means for receiving said radiant energy transmitted through said tube by said light means, and for detecting variations in intensity of said energy transmitted through said tube.
Claim 28
Dependent Claim
139 words
A liquid sensor for detecting translations between gases and liquids in a translucent tube comprising: two substantially flat substrates (217, 218) constructed of a resilient material held in a substantially opposed position relative to each other and a fixed distance apart near a first end; holding means (229) located opposite said first end and between said substrates for holding said tube, the thickness of said tube and said holding means together exceeding said fixed distance; a light means (270) held by said holder means for providing radiant energy from one side of said tube; detector means (271) held by said holder means on the opposite side of said tube from said light means for receiving said radiant energy transmitted through said tube by said light means, and for detecting variations in intensity of said energy transmitted through said tube.
Claim 29
Dependent Claim
130 words
A container (105) for holding fluid comprising: two substantially parallel sides; two other sides symmetrically located opposite each other and connected to said two substantially parallel sides to form a box-like shape; a bottom portion connected to said sides having an internal configuration of a vee-like trough, with the length of said trough perpendicular to said parallel sides, and said bottom portion having an exterior portion at the center of said trough which is substantially flat; a flange (145) connected to said bottom portion and between said parallel sides, orthogonal to the length of said trough and having a flat surface substantially coextensive with said flat surface at the center of the trough; a top portion having a septum therein and connected to said sides to form a closed volume.
Claim 30
Dependent Claim
130 words
A container (105) for holding fluid comprising: two substantially parallel sides; two other sides symmetrically located opposite each other and connected to said two substantially parallel sides to form a box-like shape; a bottom portion connected to said sides having an internal configuration of a vee-like trough, with the length of said trough perpendicular to said parallel sides, and said bottom portion having an exterior portion at the center of said trough which is substantially flat; a flange (145) connected to said bottom portion and between said parallel sides, orthogonal to the length of said trough and having a flat surface substantially coextensive with said flat surface at the center of the trough; a top portion having a septum therein and connected to said sides to form a closed volume.
Claim 31
Dependent Claim
116 words
An autodelivery system for use in an automated peptide synthesizer for delivering premeasured quantities of amino acid to said synthesizer, comprising: a track (101) for holding an array of individual containers (105) of amino acid and for directing the motion- of said cartridges; force means for pushing on said array of cartridges in a direction along the track; ejector means (113) located opposite said force means for opposing said force means, and for holding said array fixed when amino acid is being extracted from a cartridge, and for ejecting a cartridge from said array when amino acid has been extracted therefrom; extraction means (121, 123) located near said track for extracting amino acid from said cartridges.
Claim 32
Dependent Claim
116 words
An autodelivery system for use in an automated peptide synthesizer for delivering premeasured quantities of amino acid to said synthesizer, comprising: a track (101) for holding an array of individual containers (105) of amino acid and for directing the motion- of said cartridges; force means for pushing on said array of cartridges in a direction along the track; ejector means (113) located opposite said force means for opposing said force means, and for holding said array fixed when amino acid is being extracted from a cartridge, and for ejecting a cartridge from said array when amino acid has been extracted therefrom; extraction means (121, 123) located near said track for extracting amino acid from said cartridges.
References Cited (Backward Citations)
No citation records indexed.
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