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A monoclonal antibo...
A monoclonal antibody-based sandwich ELISA for measuring canine Thymidine kinase 1 protein and its role as biomarker in canine lymphoma
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- Sharif, Hanan (author)
- Alertix Vet Diagnost AB, Stockholm, Sweden.;Swedish Univ Agr Sci, Biomed Ctr, Dept Anat Physiol & Biochem, Uppsala, Sweden.
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- Saellstrom, Sara (author)
- Swedish Univ Agr Sci, Univ Anim Hosp, Uppsala, Sweden.
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- Kolli, Bhavya (author)
- Alertix Vet Diagnost AB, Stockholm, Sweden.
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- Jagarlamudi, Kiran Kumar (author)
- Alertix Vet Diagnost AB, Stockholm, Sweden.;AroCell AB, Stockholm, Sweden.
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- Wang, Liya (author)
- Swedish Univ Agr Sci, Biomed Ctr, Dept Anat Physiol & Biochem, Uppsala, Sweden.
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- Ronnberg, Henrik (author)
- Swedish Univ Agr Sci, Univ Anim Hosp, Uppsala, Sweden.;Swedish Univ Agr Sci, Dept Clin Sci, Ctr Clin Comparat Oncol C3O, Uppsala, Sweden.
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- Eriksson, Staffan (author)
- Alertix Vet Diagnost AB, Stockholm, Sweden.;Swedish Univ Agr Sci, Biomed Ctr, Dept Anat Physiol & Biochem, Uppsala, Sweden.
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Alertix Vet Diagnost AB, Stockholm, Sweden;Swedish Univ Agr Sci, Biomed Ctr, Dept Anat Physiol & Biochem, Uppsala, Sweden. Swedish Univ Agr Sci, Univ Anim Hosp, Uppsala, Sweden. (creator_code:org_t)
- Frontiers Media SA, 2023
- 2023
- English.
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In: Frontiers in Veterinary Science. - : Frontiers Media SA. - 2297-1769. ; 10
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Abstract
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- Introduction: Dogs play an important role in society, which increased during the covid epidemics. This has led to a much higher workload for the veterinarians. Therefore, there is a need for efficient diagnostic tools to identify risk of malignant diseases. Here the development of a new test that can solve some of these problems is presented. It is based on serum Thymidine Kinase 1 (TK1), which is a biomarker for cell proliferation and cell lysis.Methods: Anti-TK1 monoclonal antibodies were produced against two different epitopes, the active site of the TK1 protein and the C-terminal region of canine TK1. The antibodies were developed with hybridoma technology and validated using dot blot, Quartz Crystal Microbalance (QCM) technology, western blots, immunoprecipitation (IP), and enzyme-linked immunosorbent assay (ELISA). Clinical evaluation of Canine TK1 ELISA was done by using sera from 131 healthy dogs and 93 dogs with lymphoma. The two selected Anti-TK1 monoclonal antibodies have Kd values in the range of 10(-9) M and further analysis with dot and western blots confirmed the high affinity binding of these antibodies. A sandwich Canine TK1 ELISA was developed using the anti-TK1 antibodies, and TK1 concentrations in serum samples were determined using dog recombinant TK1 as a standard.Results: Serum TK1 protein levels were significantly higher in dogs with lymphoma compared to those in healthy dogs (p < 0.0001). Receiver operating curve analysis showed that the canine TK1-ELISA obtain a sensitivity of 0.80, at a specificity of 0.95. Moreover, the Canine TK1 ELISA has a positive predictive value (PPV) of 97%, and the negative predictive value (NPV) of 83%, reflecting the proportion of test results that are truly positive and negative. Furthermore, Canine TK1 ELISA had significantly higher capacity to differentiate dogs with T-cell lymphoma from those with B-cell lymphoma compared to earlier used TK1 activity assays.Discussion: These results demonstrate that the Canine TK1 ELISA can serve as an efficient tool in the diagnosis and management of dogs with lymphomas.
Subject headings
- LANTBRUKSVETENSKAPER -- Veterinärmedicin -- Klinisk vetenskap (hsv//swe)
- AGRICULTURAL SCIENCES -- Veterinary Science -- Clinical Science (hsv//eng)
- LANTBRUKSVETENSKAPER -- Veterinärmedicin -- Annan veterinärmedicin (hsv//swe)
- AGRICULTURAL SCIENCES -- Veterinary Science -- Other Veterinary Science (hsv//eng)
- LANTBRUKSVETENSKAPER -- Veterinärmedicin -- Medicinsk biovetenskap (hsv//swe)
- AGRICULTURAL SCIENCES -- Veterinary Science -- Medical Bioscience (hsv//eng)
Keyword
- canine TK1 ELISA
- monoclonal antibody
- immunoassay
- serum TK1 activity
- serum TK1 concentration
- canine lymphoma
- blood biomarker
- tumor marker
Publication and Content Type
- ref (subject category)
- art (subject category)
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