Functional diversification of hybridoma-produced antibodies by CRISPR/HDR genomic engineering

Johan M S van der Schoot; Felix L Fennemann; Michael Valente; Yusuf Dolen; Iris M Hagemans; Anouk M D Becker; Camille M Le Gall; Duco van Dalen; Alper Cevirgel; Jaco A C van Bruggen; Melanie Engelfriet; Tomislav Caval; Arthur E H Bentlage; Marieke F Fransen; Maaike Nederend; Jeanette H W Leusen; Albert J R Heck; Gestur Vidarsson; Carl G Figdor; Martijn Verdoes; Ferenc A Scheeren

doi:10.1126/sciadv.aaw1822

Functional diversification of hybridoma-produced antibodies by CRISPR/HDR genomic engineering

Johan M S van der Schoot, Felix L Fennemann, Michael Valente, Yusuf Dolen, Iris M Hagemans, Anouk M D Becker, Camille M Le Gall, Duco van Dalen, Alper Cevirgel, Jaco A C van Bruggen, Melanie Engelfriet, Tomislav Caval, Arthur E H Bentlage, Marieke F Fransen, Maaike Nederend, Jeanette H W Leusen, Albert J R Heck, Gestur Vidarsson, Carl G Figdor, Martijn VerdoesFerenc A Scheeren

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Abstract

Hybridoma technology is instrumental for the development of novel antibody therapeutics and diagnostics. Recent preclinical and clinical studies highlight the importance of antibody isotype for therapeutic efficacy. However, since the sequence encoding the constant domains is fixed, tuning antibody function in hybridomas has been restricted. Here, we demonstrate a versatile CRISPR/HDR platform to rapidly engineer the constant immunoglobulin domains to obtain recombinant hybridomas, which secrete antibodies in the preferred format, species, and isotype. Using this platform, we obtained recombinant hybridomas secreting Fab' fragments, isotype-switched chimeric antibodies, and Fc-silent mutants. These antibody products are stable, retain their antigen specificity, and display their intrinsic Fc-effector functions in vitro and in vivo. Furthermore, we can site-specifically attach cargo to these antibody products via chemoenzymatic modification. We believe that this versatile platform facilitates antibody engineering for the entire scientific community, empowering preclinical antibody research.

Original language	English
Article number	eaaw1822
Number of pages	12
Journal	Science advances
Volume	5
Issue number	8
DOIs	https://doi.org/10.1126/sciadv.aaw1822
Publication status	Published - 28 Aug 2019

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van der Schoot, J. M. S., Fennemann, F. L., Valente, M., Dolen, Y., Hagemans, I. M., Becker, A. M. D., Le Gall, C. M., van Dalen, D., Cevirgel, A., van Bruggen, J. A. C., Engelfriet, M., Caval, T., Bentlage, A. E. H., Fransen, M. F., Nederend, M., Leusen, J. H. W., Heck, A. J. R., Vidarsson, G., Figdor, C. G., ... Scheeren, F. A. (2019). Functional diversification of hybridoma-produced antibodies by CRISPR/HDR genomic engineering. Science advances, 5(8), Article eaaw1822. https://doi.org/10.1126/sciadv.aaw1822

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title = "Functional diversification of hybridoma-produced antibodies by CRISPR/HDR genomic engineering",

abstract = "Hybridoma technology is instrumental for the development of novel antibody therapeutics and diagnostics. Recent preclinical and clinical studies highlight the importance of antibody isotype for therapeutic efficacy. However, since the sequence encoding the constant domains is fixed, tuning antibody function in hybridomas has been restricted. Here, we demonstrate a versatile CRISPR/HDR platform to rapidly engineer the constant immunoglobulin domains to obtain recombinant hybridomas, which secrete antibodies in the preferred format, species, and isotype. Using this platform, we obtained recombinant hybridomas secreting Fab' fragments, isotype-switched chimeric antibodies, and Fc-silent mutants. These antibody products are stable, retain their antigen specificity, and display their intrinsic Fc-effector functions in vitro and in vivo. Furthermore, we can site-specifically attach cargo to these antibody products via chemoenzymatic modification. We believe that this versatile platform facilitates antibody engineering for the entire scientific community, empowering preclinical antibody research.",

author = "{van der Schoot}, {Johan M S} and Fennemann, {Felix L} and Michael Valente and Yusuf Dolen and Hagemans, {Iris M} and Becker, {Anouk M D} and {Le Gall}, {Camille M} and {van Dalen}, Duco and Alper Cevirgel and {van Bruggen}, {Jaco A C} and Melanie Engelfriet and Tomislav Caval and Bentlage, {Arthur E H} and Fransen, {Marieke F} and Maaike Nederend and Leusen, {Jeanette H W} and Heck, {Albert J R} and Gestur Vidarsson and Figdor, {Carl G} and Martijn Verdoes and Scheeren, {Ferenc A}",

note = "Publisher Copyright: Copyright {\textcopyright} 2019 The Authors, some rights reserved;",

year = "2019",

month = aug,

day = "28",

doi = "10.1126/sciadv.aaw1822",

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van der Schoot, JMS, Fennemann, FL, Valente, M, Dolen, Y, Hagemans, IM, Becker, AMD, Le Gall, CM, van Dalen, D, Cevirgel, A, van Bruggen, JAC, Engelfriet, M, Caval, T, Bentlage, AEH, Fransen, MF, Nederend, M, Leusen, JHW, Heck, AJR, Vidarsson, G, Figdor, CG, Verdoes, M & Scheeren, FA 2019, 'Functional diversification of hybridoma-produced antibodies by CRISPR/HDR genomic engineering', Science advances, vol. 5, no. 8, eaaw1822. https://doi.org/10.1126/sciadv.aaw1822

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AU - van der Schoot, Johan M S

AU - Fennemann, Felix L

AU - Valente, Michael

AU - Dolen, Yusuf

AU - Hagemans, Iris M

AU - Becker, Anouk M D

AU - Le Gall, Camille M

AU - van Dalen, Duco

AU - Cevirgel, Alper

AU - van Bruggen, Jaco A C

AU - Engelfriet, Melanie

AU - Caval, Tomislav

AU - Bentlage, Arthur E H

AU - Fransen, Marieke F

AU - Nederend, Maaike

AU - Leusen, Jeanette H W

AU - Heck, Albert J R

AU - Vidarsson, Gestur

AU - Figdor, Carl G

AU - Verdoes, Martijn

AU - Scheeren, Ferenc A

PY - 2019/8/28

Y1 - 2019/8/28

N2 - Hybridoma technology is instrumental for the development of novel antibody therapeutics and diagnostics. Recent preclinical and clinical studies highlight the importance of antibody isotype for therapeutic efficacy. However, since the sequence encoding the constant domains is fixed, tuning antibody function in hybridomas has been restricted. Here, we demonstrate a versatile CRISPR/HDR platform to rapidly engineer the constant immunoglobulin domains to obtain recombinant hybridomas, which secrete antibodies in the preferred format, species, and isotype. Using this platform, we obtained recombinant hybridomas secreting Fab' fragments, isotype-switched chimeric antibodies, and Fc-silent mutants. These antibody products are stable, retain their antigen specificity, and display their intrinsic Fc-effector functions in vitro and in vivo. Furthermore, we can site-specifically attach cargo to these antibody products via chemoenzymatic modification. We believe that this versatile platform facilitates antibody engineering for the entire scientific community, empowering preclinical antibody research.

AB - Hybridoma technology is instrumental for the development of novel antibody therapeutics and diagnostics. Recent preclinical and clinical studies highlight the importance of antibody isotype for therapeutic efficacy. However, since the sequence encoding the constant domains is fixed, tuning antibody function in hybridomas has been restricted. Here, we demonstrate a versatile CRISPR/HDR platform to rapidly engineer the constant immunoglobulin domains to obtain recombinant hybridomas, which secrete antibodies in the preferred format, species, and isotype. Using this platform, we obtained recombinant hybridomas secreting Fab' fragments, isotype-switched chimeric antibodies, and Fc-silent mutants. These antibody products are stable, retain their antigen specificity, and display their intrinsic Fc-effector functions in vitro and in vivo. Furthermore, we can site-specifically attach cargo to these antibody products via chemoenzymatic modification. We believe that this versatile platform facilitates antibody engineering for the entire scientific community, empowering preclinical antibody research.

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VL - 5

JO - Science advances

JF - Science advances

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M1 - eaaw1822

ER -

Functional diversification of hybridoma-produced antibodies by CRISPR/HDR genomic engineering

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