Construction method and application of chimeric antigen receptor (CAR) based on BMCA nano-antibody sequence

A chimeric antigen receptor and nanobody technology, applied in the field of cellular immunity, can solve the problems of long tumor killing ability and short nanobody expression sequence, and achieve the effect of strong tumor killing ability, good specificity and space saving.

Inactive Publication Date: 2019-04-26
深圳科康临床医学研究有限公司
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Problems solved by technology

[0005] Aiming at the deficiencies of the prior art, the present invention provides a method for constructing a chimeric antigen receptor based on the BMCA nanobody sequence and its application, which has a shorter expression sequence of the prepared nanobody and saves limited lentiviral vector space, The chimeric an...

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Embodiment Construction

[0022] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0023] A method for constructing a chimeric antigen receptor based on BMCA nanobody sequence, the specific steps are as follows:

[0024] Step 1: BCMA single domain antibody library construction

[0025] The alpacas were immunized regularly by expressing the extracellular region (1-54 amino acids) of BCMA protein in Escherichia coli. During immunization, blood sampling was performed to evaluate the effect of immune response. After immunization, peripheral blood ...

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Abstract

The invention relates to the technical field of cellular immunity and discloses a construction method and application of a chimeric antigen receptor (CAR) based on a BMCA nano-antibody sequence. The construction method comprises the following steps: step 1: constructing a BMCA single domain antibody library; step 2: screening a BMCA specific nano-antibody sequence; step 3: preparing chimeric antigen receptor T cells. The chimeric antigen receptor is mainly applied to tumor treatment medicines; tumors are tumor diseases related to blood and the tumor diseases related to the blood mainly comprise various leukemia and malignant lymphomas. According to the construction method and application of the chimeric antigen receptor based on the BMCA nano-antibody sequence provided by the invention, the prepared nano-antibody expression sequence is shorter and a limited lentiviral vector space is saved; the chimeric antigen receptor T cells which are constructed based on the antibody have better specificity and a stronger tumor killing capability.

Description

technical field [0001] The invention relates to the technical field of cellular immunity, in particular to a construction method and application of a chimeric antigen receptor based on BMCA nanobody sequence. Background technique [0002] In recent years, significant progress has been made in adoptive immunotherapy of tumors based on chimeric antigen receptor (CAR) modified T cells. The modified T cells not only have the ability to target tumor cells, but also overcome the local immunosuppressive microenvironment of the tumor and break the state of host immune tolerance. CAR has been developed to the fourth generation through continuous improvement. The first-generation CAR is mainly composed of scFv and ITAM (usually CD3ζ) that recognize tumor-associated antigens, but due to only CD3ζ activation signal, T cells cannot fully proliferate, and activated T cells have a short survival time in vivo. The second-generation CAR adds a costimulatory molecule such as CD28, CD134, CD...

Claims

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Application Information

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IPC IPC(8): C07K16/40C07K19/00C12N15/867C12N5/10A61K35/17A61P35/00A61P35/02
CPCA61P35/00A61P35/02A61K35/17C12N5/0636C12N15/86C07K16/005C07K16/40C07K2317/22C07K2317/569C07K2319/03C07K2319/02C07K2319/00C12N2510/00C12N2740/15043C12N2800/107
Inventor 梅志超王雨罗新高
Owner 深圳科康临床医学研究有限公司
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