Development of humanised antibodies for Crimean-Congo Haemorrhagic fever virus: Comparison of hybridoma-based versus phage library techniques
Humanised antibodies targeting Crimean-Congo Haemorrhagic virus (CCHFV) are needed for the development and standardisation of serological assays. These assays are needed to address a shortfall in available tests that meet regulatory diagnostic standards and to aid surveillance activities to extend k...
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| Published in | Journal of immunological methods Vol. 512; p. 113405 |
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| Main Authors | , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Netherlands
Elsevier B.V
01.01.2023
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| Abstract | Humanised antibodies targeting Crimean-Congo Haemorrhagic virus (CCHFV) are needed for the development and standardisation of serological assays. These assays are needed to address a shortfall in available tests that meet regulatory diagnostic standards and to aid surveillance activities to extend knowledge on the distribution of CCHFV. To generate a humanised monoclonal antibody against CCHFV, we have compared two methods: the traditional mouse hybridoma approach with subsequent sequencing and humanisation of antibodies versus a non-animal alternative using a human combinatorial antibody library (HuCAL). Our results demonstrated that the mouse hybridoma followed by humanisation protocol gave higher affinity antibodies. Whilst not yet able to demonstrate the generation of equivalent humanised antibodies without the use of animals, sequencing data enables the subsequent production of recombinant antibodies, thus providing a reduction in future animal usage for this application. Ultimately, our report provides information on development of a humanised standardised control, which can form an important positive control component of serological assays against CCHFV. |
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| AbstractList | Humanised antibodies targeting Crimean-Congo Haemorrhagic virus (CCHFV) are needed for the development and standardisation of serological assays. These assays are needed to address a shortfall in available tests that meet regulatory diagnostic standards and to aid surveillance activities to extend knowledge on the distribution of CCHFV. To generate a humanised monoclonal antibody against CCHFV, we have compared two methods: the traditional mouse hybridoma approach with subsequent sequencing and humanisation of antibodies versus a non-animal alternative using a human combinatorial antibody library (HuCAL). Our results demonstrated that the mouse hybridoma followed by humanisation protocol gave higher affinity antibodies. Whilst not yet able to demonstrate the generation of equivalent humanised antibodies without the use of animals, sequencing data enables the subsequent production of recombinant antibodies, thus providing a reduction in future animal usage for this application. Ultimately, our report provides information on development of a humanised standardised control, which can form an important positive control component of serological assays against CCHFV. |
| ArticleNumber | 113405 |
| Author | Graves, Leo P. King, Linda A. Possee, Robert D. Hewson, Roger Chambers, Adam Graham, Victoria A. Alakeely, Riyadh A. Dowall, Stuart D. Kennedy, Emma |
| Author_xml | – sequence: 1 givenname: Stuart D. surname: Dowall fullname: Dowall, Stuart D. email: stuart.dowall@ukhsa.gov.uk organization: United Kingdom Health Security Agency (UKHSA), Porton Down, Salisbury, Wiltshire SP4 0JG. UK – sequence: 2 givenname: Leo P. surname: Graves fullname: Graves, Leo P. organization: Oxford Expression Technologies, Bioinnovation Hub, Gipsy Lane, Oxford OX3 0BP. UK – sequence: 3 givenname: Emma surname: Kennedy fullname: Kennedy, Emma organization: United Kingdom Health Security Agency (UKHSA), Porton Down, Salisbury, Wiltshire SP4 0JG. UK – sequence: 4 givenname: Victoria A. surname: Graham fullname: Graham, Victoria A. organization: United Kingdom Health Security Agency (UKHSA), Porton Down, Salisbury, Wiltshire SP4 0JG. UK – sequence: 5 givenname: Riyadh A. surname: Alakeely fullname: Alakeely, Riyadh A. organization: Oxford Expression Technologies, Bioinnovation Hub, Gipsy Lane, Oxford OX3 0BP. UK – sequence: 6 givenname: Adam surname: Chambers fullname: Chambers, Adam organization: Oxford Expression Technologies, Bioinnovation Hub, Gipsy Lane, Oxford OX3 0BP. UK – sequence: 7 givenname: Robert D. surname: Possee fullname: Possee, Robert D. organization: Oxford Expression Technologies, Bioinnovation Hub, Gipsy Lane, Oxford OX3 0BP. UK – sequence: 8 givenname: Linda A. surname: King fullname: King, Linda A. organization: Department of Biological & Medical Sciences, Oxford Brookes University, Oxford OX3 0BP. UK – sequence: 9 givenname: Roger surname: Hewson fullname: Hewson, Roger organization: United Kingdom Health Security Agency (UKHSA), Porton Down, Salisbury, Wiltshire SP4 0JG. UK |
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| Keywords | Virus Hybridoma Serological Crimean-Congo HuCAL Non-animal |
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| Title | Development of humanised antibodies for Crimean-Congo Haemorrhagic fever virus: Comparison of hybridoma-based versus phage library techniques |
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