CN114544938A - Solid support comprising a set of protein arrays - Google Patents
Solid support comprising a set of protein arrays Download PDFInfo
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- CN114544938A CN114544938A CN202110820292.3A CN202110820292A CN114544938A CN 114544938 A CN114544938 A CN 114544938A CN 202110820292 A CN202110820292 A CN 202110820292A CN 114544938 A CN114544938 A CN 114544938A
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Abstract
The present invention relates to a solid support comprising a set of protein and/or peptide arrays for the determination of allergen specific antibodies, wherein said arrays are surrounded by a surrounding predetermined breaking point defining a solid support element.
Description
Technical Field
The present invention relates to the field of diagnostics, in particular allergy diagnostics.
Background
Currently available protein and peptide arrays are manufactured in a predetermined format, typically a chip containing one or more identically prepared protein and peptide arrays (see fig. 1), which then need to be processed in the laboratory by manual pipetting. A disadvantage of the known chip comprising more than one array is that the spotting of the microarray is performed directly on a relatively large chip. Examples of such disadvantageous chips and devices include ImmunOCAP ISAC (Jakob T et al Allergo J (2015)24(8):42-56) and MeDALL allergen chips (Lupinek C et al Methods (2014)66(1): 106-.
Furthermore, protein and peptide arrays are typically prepared directly on a substrate, which is then used to perform specific analytical methods. These substrates have or are typically embedded in a predetermined architecture. A specific architecture is usually required to accommodate the respective analysis device. Examples of such unfavorable chips and devices include ImmunoCAP ISAC, MeDALL allergen chips and macroarray ALEX from Thermofisher (Heffler E et al World Allergy organic J (2018)11(1):7), where ImmunoCAP ISAC and MeDALL comprise multiple arrays and macroarray ALEX comprises only one array.
Thus, depending on the equipment used, different substrates carrying protein and peptide arrays need to be used.
Another disadvantage of substrates comprising multiple protein and peptide arrays is that one poor quality array will result in the entire substrate (e.g., chip) being discarded, although other arrays present on the substrate may meet the quality criteria.
It is an object of the present invention to provide a flexible system that allows protein and peptide arrays to be mounted on multiple substrates.
Disclosure of Invention
Accordingly, the present invention relates to a solid support comprising a set of protein and/or nucleic acid peptide arrays, wherein said arrays are surrounded by a circumferential predetermined breaking point defining solid support elements.
The solid support of the invention comprises one or more, preferably at least two, arrays of proteins and/or peptides, wherein each of these arrays is separated by one or more break points surrounding these arrays. A break point around the array is required to break the element "solid support element" from a solid support comprising an array of proteins and/or peptides on its surface. These solid support elements can be placed on corresponding supports and then used for analytical purposes.
One of the main advantages of solid support elements is that they can be arranged close to each other for spotting at shorter distances than e.g. preformed on a chip. Thus, compared to pre-fabricated chips, spotter machines (e.g., microarray printers) perform shorter movements when spotting tightly packed solid support elements, which speeds up production by reducing production time.
Furthermore, the solid support of the invention comprising a set of protein and/or peptide arrays helps to keep material losses low, since individual elements of the solid support, which may be defective, can be removed without discarding the entire device, e.g. a chip, comprising the array on its surface.
One of the most important advantages of the solid support of the invention comprising a single solid support element is that after spotting the array, the single elements can be assembled in different formats for different uses. This allows, for example, the production of devices comprising only one type of array for rapid testing of a single sample, or devices comprising multiple different arrays on a solid support element for testing of different samples. Typical devices that may contain solid support elements include plates (e.g., ELISA plate format) for automated processing of samples or chips. Thus, the array printed on the solid support element allows different devices to be assembled for testing based on one standard element.
Another aspect of the invention relates to a method for producing a device comprising an array of proteins and/or peptides, comprising the steps of:
-providing a solid support according to the invention,
-breaking the circumferential predetermined breaking point of the solid support to remove solid support elements comprising the protein and/or peptide array, and
-mounting said solid support element on a chip, microarray ELISA plate or ELISA plate carrier.
The solid supports of the invention can be used to produce devices comprising solid support elements that are cleaved from the solid support. These elements may be assembled/mounted/on the device and then used to analyze the sample.
Drawings
FIG. 1 shows a chip comprising a fixed array of proteins or peptides. Such chips are known in the art.
FIG. 2 shows a solid support element 1 broken away from a solid support 3 of the present invention. The solid support element comprises an array 2 of protein and/or peptide spots immobilized/spotted on its surface.
FIG. 3 shows a solid support 3 comprising a set of protein spots and an array 2 of peptide spots. The array is surrounded by a circumferential predetermined breaking point defining solid support elements as shown in fig. 2.
FIG. 4 shows a solid support (e.g., a silicon wafer) comprising a plurality of protein and/or peptide arrays on its surface. One or more arrays may be broken or cut and assembled on various platforms (e.g., chips or boards).
Detailed Description
The solid support of the present invention comprises proteins and/or peptides spotted thereon, forming at least two, more preferably at least three, more preferably at least five, more preferably at least ten arrays. The arrays are separated from each other and surrounded by a break point. These breaking points define solid support elements that can be broken from the solid support.
As defined herein, "protein arrays" and "peptide arrays" are spatially defined arrangements of protein and peptide moieties (so-called protein spots or peptide spots) patterned on the surface of a solid support of the invention. One type of protein or peptide forms a dot. Preferably, the protein and peptide moieties are attached directly or indirectly to the surface of the solid support. Attachment may be non-specific (e.g., by physical absorption onto a surface or by formation of non-specific covalent interactions). Methods for immobilizing proteins and peptides on solid supports are well known in the art.
The number of protein spots and peptide spots attached to the solid support forming an array according to the invention is determined to be sufficient to satisfy experimental, commercial or clinical interest and may vary between 1 and 10000, preferably 1 to 1000, more preferably 1 to 500, more preferably 1 to 400, more preferably 1 to 300, more preferably 1 to 200, more preferably 1 to 100, more preferably 1 to 75, more preferably 1 to 50.
The solid support of the invention may comprise one or more protein and/or peptide arrays. It is particularly preferred that one array forms one solid support element that can be cleaved from the solid support.
As defined herein, a "circumferential predetermined breaking point" can be formed on a solid support in a variety of ways, provided that application of a force or stress to the solid support element or breaking point results in a break. For example, one or more break points can be cut through the solid support, leaving a plurality of individual solid support elements and/or contact points between the solid supports. The breaking point may be formed as a groove, e.g. by removing material by etching, laser, etc., such that the thickness of the material of the solid support at the breaking point is reduced by at least 50%, preferably at least 60%, more preferably at least 70%, more preferably at least 80%, more preferably at least 90%, more preferably at least 95% compared to the thickness of the solid support. The circumferential predetermined breaking point may also be a perforation of a solid support element surrounding the solid support. The perforations may be of any form (e.g., elongated, pierced).
The circumferential predetermined breaking point is therefore preferably a circumferential weak point, a groove, a circumferential perforation or a combination thereof of the cross section.
According to a preferred embodiment of the invention, the solid support comprises or consists of silicon, a plastic material or a metal.
The solid support may comprise or consist of various materials. In a particularly preferred embodiment, the solid support consists of silicon, wherein the solid support may be a silicon wafer. For example, silicon surfaces are 5-10 times more sensitive than glass, and also allow for the detection of low allergen-specific antibody levels, such as IgE levels, with high accuracy, and the measurement can be done with very simple and inexpensive detection equipment.
Depending on the application, the solid support may also comprise a metal surface (e.g., a gold surface). The solid support may also be composed of a body of a material (e.g., plastic) that includes a modified surface or a metal (e.g., gold) surface.
According to a preferred embodiment of the invention, the array of proteins on the solid support comprises at least two types of immobilized allergens or fragments thereof.
The allergen on the solid support may be a full-length allergen or a mature allergen. The solid support may further comprise allergen fragments, wherein these fragments preferably comprise an allergen region comprising an IgE binding motif.
According to another preferred embodiment of the invention, the allergen is selected from the group consisting of at least two allergens selected from the group consisting of: α Gal, Act d 1, Act d 2, Act d 3, Act d 4, Act d 5, Act d 6, Act d 7, Act d8, Act d 9, Act d 10, Act d 11, Act d 12, Act d 13, Aed a1, Aed a 2, Aed a 3, Aed a 4, Aed a 5, Aed a 6, Aed a 7, Aed a 8, Aed a 10, Aed a 11, Aln g1, Aln g 2, Aln g 4, Alt a1, Alt a 2, Alt a 3, Alt a 4, Alt a 5, Alt a 6, Alt a 7, Alt a 8, Alt a 9, Alt a 10, Alt a 12, Alt a13, Alt a 14, Alt a 15, Amb a1, Amb a 1.25, Amb a 1.0201, Alt a 1., Amb a 1.0202, Amb a 1.0301, Amb a 1.0302, Amb a 1.0303, Amb a 1.0304, Amb a 1.0305, Amb a 1.0401, Amb a 1.0402, Amb a 1.0501, Amb a 1.0502, Amb a 3, Amb a 4, Amb a 5, Amb a 6, Amb a 7, Amb a 8, Amb a 9, Amb a 10, Amb a 11, Amb a 12, Ana o 1, Ana o 2, Ana o 3, Ani s1, Ani s2, Ani s 3, Ani s 4, Ani s 5, Ani s 6, Ani s 7, Ani s 8, Ani s 9, Ani s 10, Ani s 11, Ani s 12, Ani s 13, Ani s 14, Api g1, Api g 2, Api 3, Api 4, Api 5, Api 6 g 5, Api s 5, Api 6, Api s 5, Api s 10, Ani s 11, Ani s 12, Ani s 13, Ani s 14, Api g1, Api g 2, Api 3, Api 5 g 6, Api 5, Api Api m 1, Api m 2, Api m 3, Api m4, Api m 5, Api m 6, Api m 7, Api m8, Api m 9, Api m 10, Api m 11, Api m 12, Api m 13kD, Ara h 1, Ara h 2, Ara h 3, Ara h 5, Ara h 6, Ara h 7, Ara h 8, Ara h 9, Ara h 10, Ara h 11, Ara h 12, Ara h 13, Ara h 14, Ara h 15, Ara h 16, Ara h 17, Art v 1, Art v 2, Art v 3, Art v 4, Art v 5, Art v 6, Asp f 1, Asp f 2, Asp f3, Asp f 4, Asp f 5, Asp f 6, Asp f 7, Asp f 8, Asp f 9, Asp f 10, Asp f 11, Asp f 12, Asp f 13, Asp f 14, Asp f 15, Asp f 16, Asp f 17, Asp f 18, Asp f 22, Asp f 23, Asp f 26, Asp f 27, Asp f 28, Asp f 29, Asp f 34, Ber e 1, Bet v 2, Bet v 3, Bet v 4, Bet v 6, Bet v 7, Bet v 8, Bla g1, Bla g 2, Bla g 3, Bla g 4, Bla g 5, Bla g 6, Bla g 7, Bla g 8, Bla g 9, Bla g 10, Bla g 11, Blot 1, Blot 2, Blot 3, Blot 4, Blot 5, Blot 6, Blot 7, Blo t 8, Blo t 9, Blo t 10, Blo t 11, Blo t 12, Blo t 13, Blo t 14, Blo t 15, Blo t 18, Blo t 19, Blo t 20, Blo t 21, Bos d 1, Bos d 2, Bos d 3, Bos d 4, Bos d 5, Bos d 6, Bos d 7, Bos d Lf, Bos d8, Bos d 9, Bos d 11, Bos d 12, aS1, aS2, b-casein, k-casein, transferrin, BSA, Bos d 6, Bran 1, Bran 4, Bra n 7, Bran 8, Can f 1, Can f 2, Can f3, Can f 4, Can f 5, Can f 6, Can f 7, Can f 8, Cas 1, Cas 2, Can f 2, Can d 4, Cas d 6, Can f 7, Can f 8, Can f 2, Cas 5, Cas 8, Cas 9, Cha o 1, Cha o 2, Cha o 3, Chea 1, Cla h 2, Cla h 5, Cla h 6, Cla h 7, Cla h 8, Cla h 9, Cla h 10, Cla h 12, Cla h 13, Cor a1, Cor a 1.0401, Cor a 2, Cor a 6, Cor a 8, Cor a 9, Cor a 10, Cor a 11, Cor a 12, Cor a13, Cor a 14, j 1, Cry j 2, Cry j 3, Cry j 4, Cyn d 1, Cup a 2, Cup a 3, Cup a 4, Cup s1, Cup s2, Cup s 3, Cyn d 1, Cyn d 2, Cyn d 4, Cyn d 5, Cyn d 6, Cyn d 7, Cyn d 11, Cyn d 12, Cyn d 13, Cyn d 15, Cyn d 22, Cyn d 23, Cyn d 24, Cyp c 1, Dau c 3, Dau c 4, Dau c 5, Der p 1, Der f 1, Der p 2, Der f 2, Der p 3, Der p 4, Der p 5, Der p 6, Der p 7, Der p 8, Der p 9, Der p 10, Der p 11, Der p 13, Der p 14, Der p 15, Der p 18, Der p 20, Der p 21, Der p 23, Clone 16, Der p 24, Deqp 36, Der p 37, Lep d 2, Equ c 1, Eu c 2, Equ c 3, Equ 4 c 4, Equ c 6, Equ c 8, Equ c 9, Equ c 10, Equ c 11, Fag e 1, Fag e 2, Fag e 3, Fag e 4, Fag e 5, Fag s1, Fag s2, Fag s 4, Fel d 1, Fel d 2, Fel d 3, Fel d 4, Fel d 5, Fel d 6, Fel d 7, Fel d8, Fra a1, Fra a 3, Fra a 4, Fra e 1, Fra 2, Fra 3, Fra e 6, Fra 7, Fra 9, Fra 10, Fra 11, Fra 12, Gad c 1, Gad m 2, Gad m 3, Gad m4, Gal d 1, Gal d 2, Gal d 3, Gal d 4, Gal d 5, Gal d 6, Gal d 7, Gal d8, Gal d 9, Gal d 10, Gly m 1, Gly m 2, Gly m 3, Gly m4, Gly m 5, Gly m 6, Gly m 7, Gly m8, Hel a1, Hel a 2, Hel a 3, Hel a 4, Hel a 6, Hev b 1, Hev b 2, Hev b 3, Hev b 4, Hev b 5, Hev b 6, Hev b 6.01, Hev b 7, Hev b 8, Hev b 9, Hev b 10, Hev b 11, Hev b 12, Hev b 13, Hev b 14, Hev b 15, Hom a1, Hom a 3, Hom a 4, Hom a 6, Hum j 1, Hum j 2, Hum j 3, Jug n 1, Jug n 2, Jug n 4, Jug r 1, Jug r 2, Jug r 3, Jug r 4, Jug r 5, Jug r 6, Jug r 7, Jug r 8, Jun a1, Jun a 2, Jun a 3, Len c 1, Len c 2, Len c 3, Lep d 2, Lep d 3, Lep d 5, Lep d 7, Lep d8, Lep d 10, Lep d 12, Lep d 13, Lep d 33, Mal d 1, Mal d 2, Mal d 3, Mal d 4, Mala s1, Mala 4, Mala 5, Mala 6, Mala 7, Mala 8, Mala 9, Mala 10, Mala 11, Mala 12, Mala 13, Mela 1, Mum 4, Mela, Mus m 7, MUXF3, Ole e 1, Ole e 2, Ole e 3, Ole e 4, Ole e 5, Ole e 6, Ole e 7, Ole e 8, Ole e 9, Ole e 10, Ole e 11, Ole e 12, Ole e 13, Ole e 14, Ole e 15, Ory c 1, Ory c 3, Ory c 4, Ory c 6, Ory c 8, Ory s1, Ory s2, Ory s 3, Ory s 7, Ory s 11, Ory s 12, Ory s 13, Ory s 14, Par j 1, Par j 2, Par j 3, Par j 4, Pen c 1, Pen c 2, Pen c 3, Pen c 6, Pen c 13, Pen c 18, Pen c 19, Pen c 22, Pen c 24, Pen c 30, Pen c 24, and Pen c 24, Pen c 32, Pen m 1, Pen m 2, Pen m 3, Pen m4, Pen m 6, Pen m8, Per a1, Per a 2, Per a 3, Per a 4, Per a 5, Per a 6, Per a 7, Per a 8, Per a 9, Per a 10, Per a 11, Per a 12, Phl p 1, Phl p 2, Phl p 3, Phl p 4, Phl p 5b, Phl p 6, Phl p 7, Phl p 11, Phl p 12, Phl p 13, Pis 1, Pis 2, Pis 3, Pis 5, Pis 6, Pis v 3, Pla a1, Pla a 2, Pla a 3, Pla a 8, Pla l 1, Pol 5, Prav 1, Pru av 2, Pru av 3, Pru av 4, Pru du 1, Pru du 2, Pru du 3, Pru du 4, Pru du 5, Pru du 6, Pru du 6.01, Pru du 6.02, Pru p 1, Pru p 2, Pru p 3, Pru p 4, Pru p 7, Que a1, Que a 2, Que a 4, Rat n 1, Rat n 4, Rat n 7, Rat n 8, Sal k 1, Sal k 2, Sal k 3, Sal k 4, Sal k 5, Sal k 6, Sal k 7, Ses i 1, Ses i 2, Ses i 3, Ses i 4, Ses i 5, Ses i 6, Ses i 7, Ses i 8, Sina 1, Sin a 2, Sin a 3, Sol a 4, Ses i 5, Ses i 6, Ses i 7, Ses i 8, Sina 1, Sin a 2, Sin a 3, Sol i 4, and s1, Sol i 2, Sol i 3, Sol i 4, Sola l 1, Sola l 2, Sola l 3, Sola l 4, Sola l 5, Sola l 6, Sola l 7, Sola t 1, Sola t 2, Sola t 3, Sola t 4, Sola t 8, Tri a1, Tri a 2, Tri a 3, Tri a 4, Tri a 5, Tri a 7, Tri a 12, Tri a13, Tri a 14, Tri a 15, Tri a 17, Tri a 18, Tri a 19, Tri a 19.0101, Tri a 20, Tri a 21, Tri a 25, Tri a 26, Tri a 27, Tri a 28, Tri a 29, Tri a 30, Tri a 31, Tri a 32, Tri a 33, Tri a 34, Tri a 35, Tri a 36, Tri a 37, Tri a 39, Tri a 40, Tri a 41, Tri a 42, Tri a 43, Tri a 44, Tri a 45, Tri a prolamin, Tri a aA _ TI, Tri a 191_369, Tri a 36, m43, m82, 10/serine, 37/Thiore, 38/GTT, 112/1-Cys, 126/Dehy, purothionin, Ves v 1, ves v 2, Ves v 3, Ves v 5, Ves v 6, Zea m 1, Zea m 2, Zea m 3, Zea m4, Zea m 5, Zea m 6, Zea m 7, Zea m8, Zea m 11, Zea m 12, Zea m 13, Zea m 14, Zea m 22, Zea m 25, Zea m 32, beta amylase, avenin and GG1, preferably selected from the group consisting of: α Gal, Act d 1, Act d 2, Act d 5, Act d8, Aln g1, Alt a 6, Amb a1, Amb a 4, Amb a 5, Amb a 6, Amb a 9, Amb a 10, Ana o 1, Ana o 2, Ana o 3, Ani s1, Ani s 3, Api g1, Api m 2, Api m4, Ara h 1, Ara h 2, Ara h 3, Ara h 6, Ara h 8, Ara h 9, Art v 1, Art v 3, Asp f 1, Asp f3, Asp f 6, Ber e 1, Bet v 2, Bet v 4, Bla g1, Bla g 2, Bla g 5, Bla g 7, Blot 5, Blas 4 Bod 4, Bos d 5, Bos d Lf, Bos d8, aS1, aS2, b-casein, k-casein, transferrin, BSA, Bos d 6, Can f 1, Can f 2, Can f3, Can f 4, Can f 5, Can f 6, Che a1, Cla h 8, Cor a 1.0401, Cor a 8, Cor a 9, Cor a 14, Cry j 1, Cyn d 1, Cup a1, Der p 1, Der f 1, Der p 2, Der f 2, Der p 4, Der p 5, Der p 7, Der p 10, Der p 11, Der p 14, Der p 15, Der p 18, Der p 21, Der p 23, Der p 37, Lep 2, Eu c 1, Equ c 3, Fel 1, Fal 1, Der p 2, Fe 2, C3, Fe 2, C1, Fe 1, C1, C1, C1, C1, C1, C1, C, Fel d 2, Fel d 4, Gad c 1, Gal d 2, Gal d 3, Gal d 5, Gly m4, Gly m 5, Gly m 6, Hev b 1, Hev b 3, Hev b 5, Hev b 6.01, Jug r 1, Jug r 2, Jug r 3, Mal d 1, Mer a1, Mus m 1, MUXF3, Ole e 1, Ole e 5, Ole e 6, Ole e 7, Ole e 8, Ole e 9, Ole e 10, Par j 2, Pen m 1, Pen m 2, Pen m4, Phl p 1, Phl p 2, Phl p 4, Phl p 5b, Phl p 6, Phl p 7, Phl p 11, Phl p 12, Pis v 3, Pla a1, Pla a 3, Pla a 3, Pla l 1, Pol d 5, Pru du 3, Pru du 4, Pru du 6, Pru du 6.01, Pru du 6.02, Pru p 1, Pru p 3, Sal k 1, Ses i 1, Tri a 14, Tri a 19.0101, Tri a aA _ TI, Tri a 191_369, Tri a 36, m43, m82, 10/serine, 37/Thiore, 38/GTT, 112/1-Cys, 126/Dehy, purothionin, Ves v 1, Ves v 5, beta-amylase, avenin and GG 1.
The solid support elements of the invention may be assembled or mounted on a device that can be used in subsequent analytical methods. In a preferred embodiment of the invention, the solid support element is used for determining the presence or amount of antibodies, in particular IgA, IgE, IgG, IgGl, IgG2, IgG3, IgG4 and IgM immunoglobulins, which bind to allergens or fragments thereof present in a sample (e.g. blood, plasma, serum, sputum). Determination of the presence of these antibodies can be used in allergy diagnosis to determine whether an individual has an allergy.
Another aspect of the invention relates to a method for producing a device comprising at least one protein and/or peptide array, comprising the steps of:
-providing a solid support according to the invention,
-breaking the circumferential predetermined breaking point of the solid support to remove solid support elements comprising the array of proteins and/or nucleic acid peptides, and
-mounting said solid support element on a chip, microarray ELISA plate or ELISA plate carrier.
One or more solid support elements can be cleaved or excised from a solid support of the invention comprising one or more, preferably more than two, protein and/or peptide arrays by applying force or using a cutting tool (e.g., a blade). Thereafter, these solid support elements are reversibly or irreversibly mounted on a device (e.g. a chip, a microarray ELISA plate or an ELISA plate carrier) for the analytical method. The reversible mounting of the solid support element is particularly advantageous as it allows the reuse of a device already mounted with the solid support element.
The solid support element is mounted to the device by affixing the solid support element to the device using glue, magnetic means, or any other mechanical means. Such securing means are well known in the art.
The present invention is further illustrated, but not limited, by the following examples.
Examples
Example 1:preparation of slide element, slide element coating and allergen sample
With 90nm silicon dioxide (SiO)2) An 8x17 mm Silicon chip was purchased from Silicon Valley Microelectronics, inc. (USA) and mounted in a frame (Ing).
Ges.m.b.h., austria). Silicon chip and glass slide (Paul Marienfeld GmbH)&Co, KG, Germany) at 0.9M (NH)4)2SO4MCP-2(Lucidant Polymers, USA) was coated at 1:100 dilution in water. After incubation of slides with MCP-2 for 15 minutes at room temperature in the dark, slides were incubated with distilled H2O rinsed and dried using a bench top centrifuge (200g,2 min). The coated slides were stored under vacuum at 4 ℃. For example, allergens were spotted in triplicate on MCP-2 coated silicon wafers and glass slides using a SciFlex array spotter (Scienion, Germany). 300pL of each allergen (c ═ 1mg/ml) were spotted at 500 μm distance from each other. 750mM Na was used 2HPO4The buffer adjusted the pH of the allergen sample to pH 8.4. After spotting, slides were incubated overnight in the dark in a 75% humidity chamber to immobilize the allergen. After overnight incubation, the chips were sprayed with blocking buffer (30 mmol/L ethanolamine in PBS containing 0.1% Tween-20) and incubated for 30 minutes at room temperature. After 30 minutes of incubation, the slides were immersed for 15 minutes in a liquid coverslipper (CANDOR Bioscience GmbH, germany). Slides were dried by centrifugation (200g,2min),and stored at 4 ℃ under vacuum until use.
Example 2:measurement of
Slides containing spotted allergens were immersed in wash buffer (PBS containing 1% Tween-20) and dried by centrifugation (200g,1min) prior to serum application. 30 microliters of undiluted (IgE assay) and diluted samples (serum and monoclonal IgE antibodies) were incubated on the microarray chip for 2 hours. For control purposes, slides were incubated with sample diluent alone (Thermo Fisher Scientific/Phadia, sweden). After incubation, slides were washed with wash buffer and dried by centrifugation (200g,1 min). Add 30 microliters with DyLightTM550-2xPEG NHS ester (Thermo Fisher Scientific, USA) conjugated mouse monoclonal anti-human IgE antibodies (Roche, Basel, Switzerland) and goat anti-human IgG antibodies (Fab') 2(Jackson ImmunoResearch, USA) (c ═ 1mg/ml) and incubated in the dark at room temperature for 30 min. Detection of antibodies and DyLight according to manufacturer's instructionsTMConjugation of 550-2xPEG NHS ester. With wash buffer and then with distillation H2O wash away unbound antibody. Slides were dried by centrifugation (200g, 2 min). Fluorescence signals were detected by a TECAN power scanner (austria) measuring 25% laser power and 50% photomultiplier tube (PMT) gain for IgE and 10% laser power and 10% PMT gain for IgG. The fluorescence intensity is analyzed, for example, using Luxscan software (China). Buffer control (i.e., results obtained with buffer and detection antibody only) was subtracted from each result and median Fluorescence Intensity (FI) from triplicate data was analyzed. Cut-off values corresponding to 0.1IU/mL for IgE and IgG detection, 100 and 10 Fluorescence Intensity (FI) for IgE and IgG, respectively.
Claims (7)
1. A solid support comprising a set of protein and/or peptide arrays for the determination of allergen-specific antibodies, wherein the arrays are surrounded by a circumferential predetermined breaking point defining a solid support element.
2. The solid support of claim 1, wherein the protein array comprises at least two immobilized allergens or fragments thereof.
3. The solid support of claim 1 or 2, wherein the allergen is selected from the group consisting of at least two allergens selected from the group consisting of: α Gal, Act d 1, Act d 2, Act d 3, Act d 4, Act d 5, Act d 6, Act d 7, Act d 8, Act d 9, Act d 10, Act d 11, Act d 12, Act d 13, Aed a 1, Aed a 2, Aed a 3, Aed a 4, Aed a 5, Aed a 6, Aed a 7, Aed a 8, Aed a 10, Aed a 11, Aln g1, Aln g 2, Aln g 4, Alt a 1, Alt a 2, Alt a 3, Alt a 4, Alt a 5, Alt a 6, Alt a 7, Alt a 8, Alt a 9, Alt a 10, Alt a 12, Alt a 13, Alt a 14, Alt a 15, Amb a 1, Amb a 1.01056, Amb a 14, Alt a 15, Amb a 1. 1.0201, Amb a 1.0202, Amb a 1.0301, Amb a 1.0302, Amb a 1.0303, Amb a 1.0304, Amb a 1.0305, Amb a 1.0401, Amb a 1.0402, Amb a 1.0501, Amb a 1.0502, Amb a 3, Amb a 4, Amb a 5, Amb a 6, Amb a 7, Amb a 8, Amb a 9, Amb a 10, Amb a 11, Amb a 12, Ana o 1, Ana o 2, Ana o 3, Ani s1, Ani s2, Ani s 3, Ani s 4, Ani s 5, Ani s 6, Ani s 7, Ani s 8, Ani s 9, Ani s 10, Ani s 11, Ani s 12, Ani s 13, Ani s 14, Api g1, Api g 2, Api 3, Api 4, Api 5, Api 6 g 5, Api s 5, Api 6, Api s 5, Api s 10, Ani s 11, Ani s 12, Ani s 13, Ani s 14, Api g1, Api g 2, Api 3, Api 5 g 6, Api 5, Api Api m 1, Api m 2, Api m 3, Api m4, Api m 5, Api m 6, Api m 7, Api m8, Api m 9, Api m 10, Api m 11, Api m 12, Api m 13kD, Ara h 1, Ara h 2, Ara h 3, Ara h 5, Ara h 6, Ara h 7, Ara h 8, Ara h 9, Ara h 10, Ara h 11, Ara h 12, Ara h 13, Ara h 14, Ara h 15, Ara h 16, Ara h 17, Art v 1, Art v 2, Art v 3, Art v 4, Art v 5, Art v 6, Asp f 1, Asp f 2, Asp f3, Asp f 4, Asp f 5, Asp f 6, Asp f 7, Asp f 8, Asp f 9, Asp f 10, Asp f 11, Asp f 12, Asp f 13, Asp f 14, Asp f 15, Asp f 16, Asp f 17, Asp f 18, Asp f 22, Asp f 23, Asp f 26, Asp f 27, Asp f 28, Asp f 29, Asp f 34, Ber e 1, Bet v 2, Bet v 3, Bet v 4, Bet v 6, Bet v 7, Bet v 8, Bla g1, Bla g 2, Bla g 3, Bla g 4, Bla g 5, Bla g 6, Bla g 7, Bla g 8, Bla g 9, Bla g 10, Bla g 11, Blot 1, Blot 2, Blot 3, Blot 4, Blot 5, Blot 6, Blot 7, Blo t 8, Blo t 9, Blo t 10, Blo t 11, Blo t 12, Blo t 13, Blo t 14, Blo t 15, Blo t 18, Blo t 19, Blo t 20, Blo t 21, Bos d 1, Bos d 2, Bos d 3, Bos d 4, Bos d 5, Bos d 6, Bos d 7, Bos d Lf, Bos d 8, Bos d 9, Bos d 11, Bos d 12, aS1, aS2, b-casein, k-casein, transferrin, BSA, Bos d 6, Bran 1, Bran 4, Bra n 7, Bran 8, Can f 1, Can f 2, Can f3, Can f 4, Can f 5, Can f 6, Can f 7, Can f 8, Cas 1, Cas 2, Can f 2, Can d 4, Cas d 6, Can f 7, Can f 8, Can f 2, Cas 5, Cas 8, Cas 9, Cha o 1, Cha o 2, Cha o 3, Chea 1, Cla h 2, Cla h 5, Cla h 6, Cla h 7, Cla h 8, Cla h 9, Cla h 10, Cla h 12, Cla h 13, Cor a 1, Cor a 1.0401, Cor a 2, Cor a 6, Cor a 8, Cor a 9, Cor a 10, Cor a 11, Cor a 12, Cor a 13, Cor a 14, j 1, Cry j 2, Cry j 3, Cry j 4, Cyn d 1, Cup a 2, Cup a 3, Cup a 4, Cup s1, Cup s2, Cup s 3, Cyn d 1, Cyn d 2, Cyn d 4, Cyn d 5, Cyn d 6, Cyn d 7, Cyn d 11, Cyn d 12, Cyn d 13, Cyn d 15, Cyn d 22, Cyn d 23, Cyn d 24, Cyp c 1, Dau c 3, Dau c 4, Dau c 5, Der p 1, Der f 1, Der p 2, Der f 2, Der p 3, Der p 4, Der p 5, Der p 6, Der p 7, Der p 8, Der p 9, Der p 10, Der p 11, Der p 13, Der p 14, Der p 15, Der p 18, Der p 20, Der p 21, Der p 23, Clone 16, Der p 24, Deqp 36, Der p 37, Lep d 2, Equ c 1, Eu c 2, Equ c 3, Equ 4 c 4, Equ c 6, Equ c 8, Equ c 9, Equ c 10, Equ c 11, Fag e 1, Fag e 2, Fag e 3, Fag e 4, Fag e 5, Fag s1, Fag s2, Fag s 4, Fel d 1, Fel d 2, Fel d 3, Fel d 4, Fel d 5, Fel d 6, Fel d 7, Fel d 8, Fra a 1, Fra a 3, Fra a 4, Fra e 1, Fra 2, Fra 3, Fra e 6, Fra 7, Fra 9, Fra 10, Fra 11, Fra 12, Gad c 1, Gad m 2, Gad m 3, Gad m4, Gal d 1, Gal d 2, Gal d 3, Gal d 4, Gal d 5, Gal d 6, Gal d 7, Gal d 8, Gal d 9, Gal d 10, Gly m 1, Gly m 2, Gly m 3, Gly m4, Gly m 5, Gly m 6, Gly m 7, Gly m8, Hel a 1, Hel a 2, Hel a 3, Hel a 4, Hel a 6, Hev b 1, Hev b 2, Hev b 3, Hev b 4, Hev b 5, Hev b 6, Hev b 6.01, Hev b 7, Hev b 8, Hev b 9, Hev b 10, Hev b 11, Hev b 12, Hev b 13, Hev b 14, Hev b 15, Hom a 1, Hom a 3, Hom a 4, Hom a 6, Hum j 1, Hum j 2, Hum j 3, Jug n 1, Jug n 2, Jug n 4, Jug r 1, Jug r 2, Jug r 3, Jug r 4, Jug r 5, Jug r 6, Jug r 7, Jug r 8, Jun a 1, Jun a 2, Jun a 3, Len c 1, Len c 2, Len c 3, Lep d 2, Lep d 3, Lep d 5, Lep d 7, Lep d 8, Lep d 10, Lep d 12, Lep d 13, Lep d 33, Mal d 1, Mal d 2, Mal d 3, Mal d 4, Mala s1, Mala 4, Mala 5, Mala 6, Mala 7, Mala 8, Mala 9, Mala 10, Mala 11, Mala 12, Mala 13, Mela 1, Mum 4, Mela, Mus m 7, MUXF3, Ole e 1, Ole e 2, Ole e 3, Ole e 4, Ole e 5, Ole e 6, Ole e 7, Ole e 8, Ole e 9, Ole e 10, Ole e 11, Ole e 12, Ole e 13, Ole e 14, Ole e 15, Ory c 1, Ory c 3, Ory c 4, Ory c 6, Ory c 8, Ory s1, Ory s2, Ory s 3, Ory s 7, Ory s 11, Ory s 12, Ory s 13, Ory s 14, Par j 1, Par j 2, Par j 3, Par j 4, Pen c 1, Pen c 2, Pen c 3, Pen c 6, Pen c 13, Pen c 18, Pen c 19, Pen c 22, Pen c 24, Pen c 30, Pen c 24, and Pen c 24, Pen c 32, Pen m 1, Pen m 2, Pen m 3, Pen m4, Pen m 6, Pen m8, Per a 1, Per a 2, Per a 3, Per a 4, Per a 5, Per a 6, Per a 7, Per a 8, Per a 9, Per a 10, Per a 11, Per a 12, Phl p 1, Phl p 2, Phl p 3, Phl p 4, Phl p 5b, Phl p 6, Phl p 7, Phl p 11, Phl p 12, Phl p 13, Pis 1, Pis 2, Pis 3, Pis 5, Pis 6, Pis v 3, Pla a 1, Pla a 2, Pla a 3, Pla a 8, Pla l 1, Pol 5, Prav 1, Pru av 2, Pru av 3, Pru av 4, Pru du 1, Pru du 2, Pru du 3, Pru du 4, Pru du 5, Pru du 6, Pru du 6.01, Pru du 6.02, Pru p 1, Pru p 2, Pru p 3, Pru p 4, Pru p 7, Que a 1, Que a 2, Que a 4, Rat n 1, Rat n 4, Rat n 7, Rat n 8, Sal k 1, Sal k 2, Sal k 3, Sal k 4, Sal k 5, Sal k 6, Sal k 7, Ses i 1, Ses i 2, Ses i 3, Ses i 4, Ses i 5, Ses i 6, Ses i 7, Ses i 8, Sina 1, Sin a 2, Sin a 3, Sol a 4, Ses i 5, Ses i 6, Ses i 7, Ses i 8, Sina 1, Sin a 2, Sin a 3, Sol i 4, and s1, Sol i 2, Sol i 3, Sol i 4, Sola l 1, Sola l 2, Sola l 3, Sola l 4, Sola l 5, Sola l 6, Sola l 7, Sola t 1, Sola t 2, Sola t 3, Sola t 4, Sola t 8, Tri a 1, Tri a 2, Tri a 3, Tri a 4, Tri a 5, Tri a 7, Tri a 12, Tri a 13, Tri a 14, Tri a 15, Tri a 17, Tri a 18, Tri a 19, Tri a 19.0101, Tri a 20, Tri a 21, Tri a 25, Tri a 26, Tri a 27, Tri a 28, Tri a 29, Tri a 30, Tri a 31, Tri a 32, Tri a 33, Tri a 34, Tri a 35, Tri a 36, Tri a 37, Tri a 39, Tri a 40, Tri a 41, Tri a 42, Tri a 43, Tri a 44, Tri a 45, Tri a prolamin, Tri a aA _ TI, Tri a 191_369, Tri a 36, m43, m82, 10/serine, 37/Thiore, 38/GTT, 112/1-Cys, 126/Dehy, purothionin, Ves v 1, ves v 2, Ves v 3, Ves v 5, Ves v 6, Zea m 1, Zea m 2, Zea m 3, Zea m4, Zea m 5, Zea m 6, Zea m 7, Zea m8, Zea m 11, Zea m 12, Zea m 13, Zea m 14, Zea m 22, Zea m 25, Zea m 32, beta amylase, avenin and GG1, preferably selected from the group consisting of: α Gal, Act d 1, Act d 2, Act d 5, Act d 8, Aln g1, Alt a 6, Amb a 1, Amb a 4, Amb a 5, Amb a 6, Amb a 9, Amb a 10, Ana o 1, Ana o 2, Ana o 3, Ani s1, Ani s 3, Api g1, Api m 2, Api m4, Ara h 1, Ara h 2, Ara h 3, Ara h 6, Ara h 8, Ara h 9, Art v 1, Art v 3, Asp f 1, Asp f3, Asp f 6, Ber e 1, Bet v 2, Bet v 4, Bla g1, Bla g 2, Bla g 5, Bla g 7, Blot 5, Blas 4 Bod 4, Bos d 5, Bos d Lf, Bos d 8, aS1, aS2, b-casein, k-casein, transferrin, BSA, Bos d 6, Can f 1, Can f 2, Can f3, Can f 4, Can f 5, Can f 6, Che a 1, Cla h 8, Cor a 1.0401, Cor a 8, Cor a 9, Cor a 14, Cry j 1, Cyn d 1, Cup a 1, Der p 1, Der f 1, Der p 2, Der f 2, Der p 4, Der p 5, Der p 7, Der p 10, Der p 11, Der p 14, Der p 15, Der p 18, Der p 21, Der p 23, Der p 37, Lep 2, Eu c 1, Equ c 3, Fel 1, Fal 1, Der p 2, Fe 2, C3, Fe 2, C1, Fe 1, C1, C1, C1, C1, C1, C1, C, Fel d 2, Fel d 4, Gad c 1, Gal d 2, Gal d 3, Gal d 5, Gly m4, Gly m 5, Gly m 6, Hev b 1, Hev b 3, Hev b 5, Hev b 6.01, Jug r 1, Jug r 2, Jug r 3, Mal d 1, Mer a 1, Mus m 1, MUXF3, Ole e 1, Ole e 5, Ole e 6, Ole e 7, Ole e 8, Ole e 9, Ole e 10, Par j 2, Pen m 1, Pen m 2, Pen m4, Phl p 1, Phl p 2, Phl p 4, Phl p 5b, Phl p 6, Phl p 7, Phl p 11, Phl p 12, Pis v 3, Pla a 1, Pla a 3, Pla a 3, Pla l 1, Pol d 5, Pru du 3, Pru du 4, Pru du 6, Pru du 6.01, Pru du 6.02, Pru p 1, Pru p 3, Sal k 1, Ses i 1, Tri a 14, Tri a 19.0101, Tri a aA _ TI, Tri a 191_369, Tri a 36, m43, m82, 10/serine, 37/Thiore, 38/GTT, 112/1-Cys, 126/Dehy, purothionin, Ves v 1, Ves v 5, beta-amylase, avenin and GG 1.
4. The solid support of any one of claims 1 to 3, wherein the solid support comprises or consists of silicon, a plastic material, or a metal.
5. The solid support according to any one of claims 1 to 4, wherein the solid support is a silicon wafer.
6. The solid support of any one of claims 1 to 6, wherein the encircling predetermined breaking point is a encircling weak point of a cross-section, a groove, an encircling perforation, or a combination thereof.
7. Method for producing a device comprising an array of proteins and/or peptides, comprising the following steps
-providing a solid support according to any one of claims 1 to 6,
-breaking the circumferential predetermined breaking point of the solid support to remove solid support elements comprising an array of proteins and/or peptides, and
-mounting said solid support element on a chip, microarray ELISA plate or ELISA plate carrier.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ATA50977/2020A AT524623B1 (en) | 2020-11-11 | 2020-11-11 | Solid support comprising a set of protein arrays |
| ATA50977/2020 | 2020-11-11 |
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| CN114544938A true CN114544938A (en) | 2022-05-27 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202110820292.3A Pending CN114544938A (en) | 2020-11-11 | 2021-07-20 | Solid support comprising a set of protein arrays |
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|---|---|
| CN (1) | CN114544938A (en) |
| AT (1) | AT524623B1 (en) |
| WO (1) | WO2022099343A1 (en) |
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|---|---|---|---|---|
| DE3045199A1 (en) * | 1980-12-01 | 1982-07-01 | Eppendorf Gerätebau Netheler + Hinz GmbH, 2000 Hamburg | Solid-phase immunoassay method - using reagent plate with perforated wells fitting into wells in sample plate |
| DE4322180A1 (en) * | 1993-07-03 | 1995-01-12 | Merck Patent Gmbh | Perforated films for thin-layer chromatography |
| DE19609315C2 (en) * | 1996-03-09 | 1998-11-05 | Univ Dresden Tech | Nasal sampler |
| DE29923907U1 (en) * | 1999-12-01 | 2001-07-26 | Quantifoil Micro Tools GmbH, 07745 Jena | Miniaturized slide for carrying out a large number of test series in the submicroliter range |
| DE10242066B4 (en) * | 2002-09-11 | 2005-03-17 | Cytopharm Gmbh | Process for the treatment of cell cultures |
| US7390457B2 (en) * | 2002-10-31 | 2008-06-24 | Agilent Technologies, Inc. | Integrated microfluidic array device |
-
2020
- 2020-11-11 AT ATA50977/2020A patent/AT524623B1/en active
-
2021
- 2021-07-20 CN CN202110820292.3A patent/CN114544938A/en active Pending
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| Publication number | Publication date |
|---|---|
| AT524623B1 (en) | 2023-02-15 |
| AT524623A1 (en) | 2022-05-15 |
| WO2022099343A1 (en) | 2022-05-19 |
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