Kit for detecting alpha-interferon and related application

Abstract

The invention discloses a kit for detecting alpha-interferon and related application, and relates to the field of biology, the kit comprises a capture protein for capturing the alpha-interferon and a detection protein for detecting the alpha-interferon; the capture protein comprises receptor protein of alpha-interferon; the detection protein comprises an anti-alpha-interferon antibody; the kit is applied to IFN-alpha detection, can cover various subtypes of IFN-alpha, including natural interferon and recombinant interferon, has no cross reaction with INF-beta and INF-epsilon, is high in detection accuracy and good in repeatability, and can quickly and effectively detect the content of IFN-alpha in human serum, so that the overall level and the active state of IFN-alpha in vivo are reflected more accurately; the method is of great significance to understanding the human immune state, diagnosing and monitoring the development of diseases and evaluating the effectiveness of treatment.

Description

Technical Field

[0001] The present invention relates to the biological field, and in particular to a kit for detecting alpha-interferon and related applications. Background Art

[0002] Interferon-α (IFN-α) is a type of cytokine with antiviral and immunomodulatory functions, which is mainly produced by macrophages, dendritic cells and epithelial cells under the induction of viruses and nucleic acids. IFN-α has a direct antiviral effect, and further exerts an indirect antiviral effect by regulating primary and secondary immune responses. Studies have found that the level of IFN-α in patients with chronic hepatitis B is significantly higher than that in healthy people. In addition, the level of IFN-α in the blood is increased in certain autoimmune diseases (such as systemic lupus erythematosus). In diseases such as multiple sclerosis, rheumatoid arthritis and inflammatory bowel disease, IFN-α has potential anti-inflammatory and tissue protective effects. In vitro quantitative detection of the concentration of IFN-α in human serum or plasma is mainly used clinically to monitor the body's immune status and inflammatory response.

[0003] IFN-α has been studied in detail and widely recognized to have 15 subtypes: IFNA-1α, IFNA-1β, IFNA-2, IFNA-2β, IFNA-4A, IFNA-4B, IFNA-5, IFNA-6, IFNA-7, IFNA-8, IFNA-10, IFNA-14, IFNA-16, IFNA-17 and IFNA-21. Each subtype has a molecular weight of 19-26kD, and is composed of 149 to 172 amino acids. It has similar sequences and structures, and the homology is close to more than 70%. However, it is found in clinical practice that the subtypes of IFN-α induced by the same virus in different cells or different viruses infecting the same cells are very different. The IFN-α chemiluminescence detection kits currently available on the market can generally only detect IFN-α1 or IFN-α2 subunits, and the flow fluorescence platform generally has problems such as low sensitivity, poor specificity, and poor repeatability. Therefore, problems such as missed detection or poor repeatability often occur in clinical testing.

[0004] In view of this, the present invention is proposed. Summary of the invention

[0005] The purpose of the present invention is to provide a kit for detecting α-interferon and related applications. In view of the problem that the prior art cannot detect all subtypes of IFN-α, the present application provides a new composition that can cover all subtypes of IFN-α (including IFNA-1α, IFNA-1β, IFNA-2, IFNA-2β, IFNA-4A, IFNA-4B, IFNA-5, IFNA-6, IFNA-7, IFNA-8, IFNA-10, IFNA-14, IFNA-16, IFNA-17 and IFNA-21), and has the advantages of high detection sensitivity and good repeatability.

[0006] The present invention is achieved in that:

[0007] In a first aspect, the present invention provides a composition for detecting IFN-α, comprising: a capture protein for capturing α-interferon and a detection protein for detecting α-interferon; wherein the capture protein comprises: a receptor protein for α-interferon, and the receptor protein for α-interferon comprises any one or both of IFNAR1 and IFNAR2; and the detection protein comprises: an anti-α-interferon antibody or an antigen-binding fragment thereof.

[0008] In some embodiments, the amino acid sequence of IFNAR1 is at least 80% identical to the sequence shown in SEQ ID NO:1.

[0009] In some embodiments, the at least 80% identity refers to any one of 80%, 82%, 84%, 86%, 88%, 90%, 92%, 94%, 96%, 98%, 100%, or any two thereof. Identity refers to the nucleotide or amino acid residues of two sequences being exactly the same at the same site.

[0010] In some embodiments, the amino acid sequence of IFNAR1 is as shown in SEQ ID NO:1.

[0011] The amino acid sequence of IFNAR2 is at least 80% identical to the sequence shown in SEQ ID NO:2.

[0012] In some embodiments, the amino acid sequence of IFNAR2 is as shown in SEQ ID NO:2.

[0013] In some embodiments, when the receptor protein of α-interferon includes a combination of IFNAR1 and IFNAR2, the mass ratio of IFNAR1 to IFNAR2 is 1 to 4: 2. The mass ratio of IFNAR1 to IFNAR2 can be any one of 1:2, 1.5:2, 2:2, 2.5:2, 3:2, 3.5:2, 4:2, or any two thereof.

[0014] In some embodiments, the mass ratio of the IFNAR1 to the IFNAR2 is 1 to 2:1.

[0015] In some embodiments, the composition further comprises: a solid phase carrier.

[0016] In some embodiments, the capture protein is coated on the solid support.

[0017] In some embodiments, the solid phase carrier includes any one or more of: magnetic beads, plates, and membranes.

[0018] In some embodiments, the ratio of capture protein to solid phase carrier is 10-50 μg / mg, specifically any one of 10, 15, 20, 25, 30, 35, 40, 45, 50 μg / mg or a range between any two thereof.

[0019] In some embodiments, the detection protein includes any one or more of: anti-IFN-α antibody of Santa Cruz Biotechnology and anti-IFN-α antibody of Mabtech.

[0020] In some embodiments, the composition further comprises a marker.

[0021] In some embodiments, the label is coupled to the detection protein.

[0022] In some embodiments, the label comprises any one or more of fluorescent dyes / proteins, enzymes, radioactive isotopes, chemiluminescent agents and nanoparticle labels.

[0023] In some embodiments, the enzyme comprises any one or more of horseradish peroxidase, alkaline phosphatase and β-galactosidase.

[0024] In some embodiments, the working concentration of the detection protein coupled to the marker is 0.1-0.6 μg / mL, specifically any one of 0.1, 0.2, 0.3, 0.4, 0.5 and 0.6 μg / mL or a range between any two thereof.

[0025] In some embodiments, the working concentration of the detection protein coupled with the marker is 0.3-0.5 μg / mL, specifically any one of 0.3, 0.4, 0.5 μg / mL or a range between any two of them.

[0026] In a second aspect, the present invention provides use of the composition described in any of the aforementioned embodiments in the preparation of an α-interferon detection kit.

[0027] In a third aspect, the present invention provides an α-interferon detection kit, comprising: the composition described in any of the aforementioned embodiments.

[0028] In some embodiments, the compositions are stored in corresponding storage solutions respectively;

[0029] For example, after the captured protein and coated magnetic beads are stored in a magnetic bead storage solution, the magnetic bead storage solution usually contains components such as a buffer, a protective protein, a surfactant, and a preservative;

[0030] After the test protein is labeled, it is stored in a labeling preservation solution. Taking alkaline phosphatase labeling as an example, the preservation solution usually contains components such as buffer, salt, reducing sugar, protective protein, and preservatives.

[0031] In some embodiments, the kit further comprises: any one or more of a standard, a dilution buffer, a coating buffer, a blocking solution, and a washing solution.

[0032] The kit of the present invention can be used to detect multiple α-interferon subtypes; the α-interferon subtypes include but are not limited to: any one or more combinations of IFNA-1α, IFNA-1β, IFNA-2, IFNA-2β, IFNA-4A, IFNA-4B, IFNA-5, IFNA-6, IFNA-7, IFNA-8, IFNA-10, IFNA-14, IFNA-16, IFNA-17 and IFNA-21.

[0033] The present invention has the following beneficial effects:

[0034] The composition provided by the present invention is used for IFN-α detection, can cover multiple subtypes of IFN-α, including natural interferon and recombinant interferon, and has no cross reaction with INF-β and INF-ε. The prepared IFN-α detection kit has high accuracy and good repeatability, and can quickly and effectively detect the IFN-α content in human serum, thereby more accurately reflecting the overall level and activity state of IFN-α in the body, which is of great significance for understanding the human immune state, diagnosing and monitoring the development of diseases, and evaluating the effectiveness of treatment. DETAILED DESCRIPTION

[0035] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0036] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0037] Example 1. Material preparation

[0038] Anti-IFN-α antibody 1 (purchased from Santa Cruz Biotechnology, catalog number sc-80996), anti-IFN-α antibody 2 (purchased from Mabtech, catalog number 3425-3-1000), alkaline phosphatase (purchased from BBI Solutions), interferon receptor protein α (IFNAR1) (purchased from R&D Systems), interferon receptor protein β (IFNAR2) (purchased from Sinobiological) are all commercially available materials.

[0039] The interferon receptor protein α (IFNAR1) sequence is as follows (SEQ ID NO: 1):

[0040] GGKNLKSPQKVEVDIIDDNFILRWNRSDESVGNVTFSFDYQKTGMDNWIKLSGCQNITSTKCNFSSLKLNVYEEIKLRIRAEKENTSSWYEVDSFTPFRKAQI GPPEVHLEAEDKAIVIHISPGTKDSVMWALDGLSFTYSLVIWKNSSGVEERIENIYSRHKIYKLSPETTYCLKVKAALLTSWKIGVYSPVHCIKTTVENELPP PENIEVSVQNQNYVLKWDYTYANMTFQVQWLHAFLKRNPGNHLYKWKQIPDCENVKTTQCVFPQNVFQKGIYLLRVQASDGNNTSFWSEEIKFDTEIQAFLLP PVFNIRSLSDSFHIYIGAPKQSGNTPVIQDYPLIYEIIFWENTSNAERKIIEKKTDVTVPNLKPLTVYCVKARAHTMDEKLNKSSVFSDAVCEKTKPGNTSK.

[0041] The interferon receptor protein β (IFNAR2) sequence is as follows (SEQ ID NO: 2):

[0042] MLLSQNAFIFRSLNLVLMVYISLVFGISYDSPDYTDESCTFKISLRNFRSILSWELKNHSIVPTHYTLLYTIMSKPEDLKVVKNCANTTRSFCDLTDEWRSTHEAYVTVLEGFSGNTTLFSC SHNFWLAIDMSFEPPFEIVGFTNHINVMVKFPSIVEEELQFDLSLVIEEQSEGIVKKHKPEIKGNMSGNFTYIIDKLIPNTNYCVSVYLEHSDEQAVIKSPLKCTLLPPGQESESAESAK.

[0043] (1) Preparation of sulfonated magnetic particle working solution

[0044] 1. Remove the supernatant from the sulfonated magnetic particles by magnetic separation, wash with PBS at pH 7.0 and concentration of 20 mM, and then dry the washing solution on a magnetic rack;

[0045] 2. Mix the antibody / protein, magnetic beads and ammonium sulfate in proportion and place on a multi-purpose rotary shaker at 4°C for 24 hours at a speed of 30 times / min;

[0046] 3. Place the centrifuge tube in step 2 on the magnetic rack for 5 minutes, discard the supernatant, wash with PBS with a pH of 7.0 and a concentration of 20mM, and aspirate the washing solution;

[0047] 4. Resuspend the washed magnetic beads in step 3 with PBS (pH 7.0, concentration 20mM), 5% BSA blocking solution, mix well and place on a multi-purpose rotary shaker at 37°C for 5 hours;

[0048] 5. Repeat step 4;

[0049] 6. Resuspend the beads in magnetic bead working solution to obtain a concentration of 0.3 mg / mL magnetic bead working solution, wherein the buffer contains 0.05 mg / mL bovine serum albumin, 0.005 mg / mL Triton X-100 and 0.005 mg / mL proclin 300, and a PBS buffer with a pH of 7.0 and a concentration of 0.1 mol / L.

[0050] (2) Preparation of alkaline phosphatase-labeled antibody / protein working solution

[0051] Sulfo-SMCC, Chinese name 4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid sulfonic acid succinimide ester sodium salt, is a water-soluble amine-thiol crosslinker.

[0052] The antibody / protein intermediate and alkaline phosphatase intermediate were prepared at a molar ratio of sulfo-SMCC: antibody / protein of 20:1 and a molar ratio of reducing agent: alkaline phosphatase of 20:1, respectively; then, the mixture was incubated at 30°C for 2h at a mass ratio of antibody / protein intermediate: alkaline phosphatase intermediate of 2:1; and alkaline phosphatase-labeled antibody / protein master solutions were prepared after further desalting.

[0053] The alkaline phosphatase-labeled antibody / protein working solution was further diluted with enzyme-labeled diluent to prepare 0.25 μg / mL. The enzyme-labeled diluent contains phosphate buffer with a pH of 6.5 and a concentration of 0.1 mol / L, 0.2 mg / mL bovine serum albumin, 0.1 mg / mL sodium chloride, 0.1 mg / ml sucrose, and 0.005 mg / mL proclin 300.

[0054] (3) Preparation of test sample working solution

[0055] The test samples used are as follows:

[0056] S: Multi-subtype natural antigen, a mixture of multi-subtype natural interferons produced by human leukocytes—Interferon alpha (Human, leukocyte derived) WHO International Standard (NIBSC, 94 / 784);

[0057] R: multi-subtype recombinant antigen, multi-subtype recombinant interferon mixture (15 recombinant IFN-α subtype proteins)—Human IFN-α All Subtype Standard (R&D Systems, PART#899172);

[0058] Cross-reactive sample 1: IFN-β, (purchased from GenScript);

[0059] Cross-reactive sample 2: IFN-ε, (purchased from MedChemExpress);

[0060] Use sample diluent to dilute each sample to the working concentration:

[0061] Multi-subtype natural antigens S1, S2 (10pg / mL, 10000pg / mL);

[0062] Multi-subtype recombinant antigens R1, R2 (10pg / mL, 10000pg / mL);

[0063] Cross-reactive samples were diluted to 10,000 pg / mL;

[0064] The sample diluent comprises: pH 7.0, phosphate buffer with a concentration of 0.01 mol / L, 0.05 mg / mL bovine serum albumin, 0.05 mg / mL sodium chloride and 0.01 mg / mL Tween20.

[0065] Example 2: Effects of different combinations of capture proteins and detection proteins on detection results

[0066] This embodiment provides a plurality of combinations of capture proteins and detection proteins formed by different proteins, and detects the detection results of different antigens.

[0067] According to the preparation method of Example 1, anti-IFN-α antibody 1 magnetic bead working solution, anti-IFN-α antibody 2 magnetic bead working solution, IFNAR1 magnetic bead working solution, and IFNAR2 magnetic bead working solution with a concentration of 0.3 mg / mL were prepared respectively. And four alkaline phosphatase-labeled antibody / protein working solutions with a concentration of 0.25 μg / mL were prepared: alkaline phosphatase-labeled anti-IFN-α antibody 1 working solution, alkaline phosphatase-labeled anti-IFN-α antibody 2 working solution, alkaline phosphatase-labeled IFNAR1 working solution, and alkaline phosphatase-labeled IFNAR2 working solution.

[0068] Different combinations of magnetic bead working solutions and alkaline phosphatase-labeled protein working solutions were used to test the samples to be tested. The combinations are shown in the table below.

[0069] Table 1 Combination

[0070]

[0071]

[0072] The combination of Table 1 was used to detect sample diluent S0 (0 pg / mL, containing no antigen), sample solutions S1 and S2 (10 pg / mL and 10000 pg / mL) diluted with a multi-subtype natural interferon mixture, and sample solutions R1 and R2 (10 pg / mL and 10000 pg / mL) diluted with a multi-subtype recombinant interferon mixture, cross-reactive sample 1 (10000 pg / mL of IFN-β), and cross-reactive sample 2 (10000 pg / mL of IFN-ε), and the test results are shown in Table 2 below.

[0073] Table 2 Test results

[0074]

[0075]

[0076] As shown in Table 2, the repeatability of Group 1 and Group 2 (when anti-IFN-α antibody 1 and anti-IFN-α antibody 2 are paired with each other) is poor (CV>3%), but there is no cross-interference to IFN-β and IFN-ε, indicating that anti-IFN-α antibody 1 and anti-IFN-α antibody 2 can recognize one or several subtypes of IFN-α, but during the antibody coating or labeling process, the antibody activity is affected or some sites are shielded, resulting in randomness in the recognition of natural or recombinant multi-subtype antigens, thereby deteriorating the repeatability.

[0077] Groups 3 and 4 (IFNRA1 and IFNRA2 are receptor proteins for INF-α, and the two receptors are paired with each other) have good repeatability with CV < 3%, indicating that the combination can recognize multiple subtypes of antigens; however, the overall signal-to-noise ratio is low, and there is a problem of cross-interference between IFN-β and IFN-ε;

[0078] Groups 5 to 8 (combination of receptor protein and antibody) had good repeatability with CV < 3%, excellent signal-to-noise ratio, and no cross-interference between IFN-β and IFN-ε. Among them, Group 5 (IFNAR1 combined with anti-IFN-α antibody 1) had the best signal-to-noise ratio, with CV within 2%, and no cross-reaction.

[0079] Example 3: Effects of different capture proteins on detection results

[0080] IFNAR1 and IFNAR2 were mixed according to the ratio in Table 3 (mass ratio, IFNAR1 and IFNAR2 were mixed and coated with magnetic beads, total protein: magnetic beads mass = 20 μg / mg), and the corresponding magnetic beads working solution was prepared by referring to the preparation method of sulfonated magnetic microparticle working solution in Example 1, and matched with alkaline phosphatase labeled anti-IFN-α antibody 1 working solution to detect the detection results of different samples. The combination of capture protein and detection protein is shown in Table 3 below.

[0081] Table 3 Combinations of capture proteins and detection proteins

[0082]

[0083] The combination of Table 3 was used to detect sample diluent S0 (0 pg / mL, containing no antigen), sample solutions S1 and S2 (10 pg / mL and 10000 pg / mL) diluted with a multi-subtype natural interferon mixture, and sample solutions R1 and R2 (10 pg / mL and 10000 pg / mL) diluted with a multi-subtype recombinant interferon mixture, cross-reaction sample 1 (10000 pg / mL of IFN-β), and cross-reaction sample 2 (10000 pg / mL of IFN-ε), and the test results are shown in Table 4 below.

[0084] Table 4 Test results

[0085]

[0086] As can be seen from Table 4, the repeatability of detection in groups 9 to 13 was good (CV < 3%), the signal-to-noise ratio was good, and there was no cross-interference between IFN-β and IFN-ε; there was little difference between group 13 and group 5, and groups 9 to 12 were improved compared with group 5 of Example 1; among them, group 10 (when IFNAR1:IFNAR2 in total protein was 1.5:1) had the best CV and specificity, and the signal-to-noise ratio was significantly improved.

[0087] Example 4: Effect of the concentration of enzyme-labeled antibody on the test results

[0088] Using the capture protein and detection protein combination of Group 10 in Example 3, the magnetic bead working solution remained unchanged, and the alkaline phosphatase-labeled anti-IFN-α antibody 1 working solutions of different concentrations were prepared according to the preparation method of the alkaline phosphatase-labeled antibody / protein working solution in Example 1, and the detection results of different samples were examined, as shown in Table 5 below.

[0089] Table 5 Combination

[0090]

[0091] The antibody combination in Table 5 was used to detect gradient reference materials P1-P5, with corresponding concentrations of 0 pg / mL, 10 pg / mL, 100 pg / mL, 1000 pg / mL, and 10000 pg / mL (obtained by diluting multi-subtype natural antigen S using antigen diluent). The test results are shown in Table 6 below.

[0092] Table 6 Test results

[0093]

[0094] The results of groups 14 to 18 showed that the repeatability and signal-to-noise ratio of each group were good; when the enzyme label concentration was 0.3-0.5μg / mL, the CV and signal-to-noise ratio were good; among them, group 16 (enzyme label concentration was 0.4μg / mL) was the best.

[0095] Example 5: Clinical application

[0096] The magnetic bead labeling working solution and the enzyme labeling working solution prepared by Group 16 in Example 4 were used in combination with the cleaning solution, substrate solution, and sample diluent produced by our company to form the α-interferon detection kit of this application. At the same time, two commercial kits, α-interferon detection kit 1 (single subtype IFN-α2) and α-interferon detection kit 2 (multi-subtype All Subtype, referring to a multi-subtype natural interferon mixture produced by human leukocytes, mainly IFNA-1α, IFNA-1β, IFNA-2, IFNA-2β, IFNA-4A, IFNA-4B, IFNA-5, IFNA-6, IFNA-7, IFNA-8, IFNA-10, IFNA-14, IFNA-16, IFNA-17, IFNA-21) were used as controls, and 500 healthy human samples, 300 systemic lupus erythematosus (SLE) patient samples, and 100 new coronavirus-positive patient samples were simultaneously detected. The test results are shown in Table 7 below.

[0097] Table 7 Test results

[0098]

[0099] Negative detection rate = number of samples with negative test results / total number of samples;

[0100] Positive detection rate = number of samples with positive test results / total number of samples.

[0101] From the results in Table 7, the sensitivity of the α-interferon detection kit prepared in this application in SLE patient samples and new coronavirus-positive patient samples is comparable to that of the commercial α-interferon detection kit 2 (multi-subtypes), reaching 34.33% and 36%, respectively, which is significantly higher than the commercial α-interferon detection kit 1 (single subtype). The test results of healthy people show that the α-interferon detection kit prepared in the examples of this application has better specificity, which is 0.6% and 1.6% higher than the commercial α-interferon detection kit 1 (single subtype) and the commercial α-interferon detection kit 2 (multi-subtypes), respectively.

[0102] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A composition for detecting interferon-α, characterized in that: It includes: A capture protein for capturing α-interferon and a detection protein for detecting α-interferon; wherein the capture protein includes: a receptor protein for α-interferon, and the receptor protein for α-interferon includes any one or both of IFNAR1 and IFNAR2; and the detection protein includes: an antibody against α-interferon or an antigen-binding fragment thereof.

2. The composition according to claim 1, characterized in that The amino acid sequence of IFNAR1 has at least 80% identity with the sequence shown in SEQ ID NO:1; The amino acid sequence of IFNAR2 has at least 80% identity with the sequence shown in SEQ ID NO:2; Optionally, the amino acid sequence of IFNAR1 is shown in SEQ ID NO: 1; Optionally, the amino acid sequence of IFNAR2 is as shown in SEQ ID NO:

2.

3. The composition according to claim 2, characterized in that The receptor protein of the α-interferon includes IFNAR1 and IFNAR2, and the mass ratio of the IFNAR1 to the IFNAR2 is 1 to 4:2; Optionally, the mass ratio is 1 to 2:

1.

4. The composition according to any one of claims 1 to 3, characterized in that The composition further comprises: a solid phase carrier; Optionally, the capture protein is coated on the solid phase carrier; Optionally, the solid phase carrier includes any one or more of magnetic beads, plates and membranes.

5. The composition according to claim 1, characterized in that The detection protein includes: any one or more of the anti-IFN-α antibody of Santa Cruz Biotechnology and the anti-IFN-α antibody of Mabtech.

6. The composition according to claim 5, characterized in that The composition further comprises a marker; Optionally, the marker is coupled to the detection protein; Optionally, the marker includes: any one or more of fluorescent dyes, fluorescent proteins, enzymes, radioactive isotopes, chemiluminescent agents and nanoparticle markers; Optionally, the enzyme includes: any one or more of horseradish peroxidase, alkaline phosphatase and β-galactosidase; Optionally, the working concentration of the detection protein coupled with the marker is 0.1 to 0.6 μg / mL; Optionally, the working concentration of the detection protein coupled with the marker is 0.3-0.5 μg / mL.

7. Use of the composition according to any one of claims 1 to 6 in the preparation of an interferon-α detection kit.

8. An α-interferon detection kit, characterized in that: It includes: The composition according to any one of claims 1 to 6.

9. The kit according to claim 8, characterized in that The kit further comprises: any one or more of a standard substance, a dilution buffer, a coating buffer, a blocking solution and a washing solution.

10. The kit according to claim 9, characterized in that The kit is capable of detecting multiple subtypes of α-interferon; Optionally, the α-interferon subtype includes any one or more combinations of IFNA-1α, IFNA-1β, IFNA-2, IFNA-2β, IFNA-4A, IFNA-4B, IFNA-5, IFNA-6, IFNA-7, IFNA-8, IFNA-10, IFNA-14, IFNA-16, IFNA-17 and IFNA-21.

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