Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Antibody producing non-human mammals

a technology for antibody production and mammals, applied in the field of production and use of non-human animals, can solve the problems of reducing the efficacy of subsequent administrations, and achieve the effects of reducing the number of administrations, facilitating the opening and maintenance of euchromatin, and preventing silencing

Inactive Publication Date: 2010-03-18
MERUS NV
View PDF0 Cites 160 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0018]A means for rendering a gene encoding an immunoglobulin chain (or chains) resistant to DNA rearrangement and / or mutation is of course removal of all genetic elements responsible for the rearrangement and / or mutation. The drawback thereof is that the variability of the two chains is eliminated, whereas the invention preferably retains the variability in one chain (preferably the heavy chain) and inhibits and / or prevents the rearrangement-mutation of the other chain (preferably the light chain).
[0071]Furthermore, it is shown that upon immunization, the murine and human VH regions in the afore-mentioned transgenic mice but not the VL regions are capable of undergoing somatic hypermutations to generate high affinity antibodies. Advantageously, these antibodies encoded by germline VL regions are predicted to contribute to lower immunogenicity when applied in humans and result in more stable antibodies that are less prone to aggregation and thus safer for therapeutic use in humans.

Problems solved by technology

One of the major impediments facing the development of in vivo applications for mAbs in humans is the intrinsic immunogenicity of non-human Igs.
Patients respond to therapeutic doses of mouse mAbs by making antibodies against the mouse Ig sequences (Human Anti Mouse Antibodies; HAMA), causing acute toxicity, alter their biodistribution and accelerate clearance, thus reducing the efficacy of subsequent administrations (Mirick et al.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Antibody producing non-human mammals
  • Antibody producing non-human mammals
  • Antibody producing non-human mammals

Examples

Experimental program
Comparison scheme
Effect test

example 1

Human Light Chain V-Gene Clones

[0107]This example describes the rationale behind the choice of two human light chain V-genes, one gene of the kappa type and one gene of the lambda type, that are used as a proof of concept for light chain expressing transgenic mice. De Wildt et al. 1999 (de Wildt et al. (1999), J. Mol. Biol. 285(3):895) analyzed the expression of human light chains in peripheral IgG-positive B-cells. Based on these data, IGKV1-39 (O12) and IGLV2-14 (2a2) were chosen as light chains as they were well represented in the B-cell repertoire. The J-segment sequence of the light chains has been chosen based upon sequences as presented in GenBank ABA26122 for IGKV1-39 (B. J. Rabquer, S. L. Smithson, A. K. Shriner and M. A. J. Westerink) and GenBank AAF20450 for IGLV2-14 (O. Ignatovich, I. M. Tomlinson, A. V. Popov, M. Bruggemann and G. J. Winter, J. Mol. Biol. 294 (2):457-465 (1999)).

[0108]All framework segments are converted into germline amino acid sequences to provide the...

example 2

Obtaining Mouse Heavy Chain V-Genes that Pair with Human IGKV1-39 Gene Segment to Form Functional Antibody Binding Sites

[0109]This example describes the identification of mouse heavy chain V-genes that are capable of pairing with a single, rearranged human germline IGKV1-39 / J region. A spleen VH repertoire from mice that were immunized with tetanus toxoid was cloned in a phage display Fab vector with a single human IGKV1-39-C kappa light chain and subjected to panning against tetanus toxoid. Clones obtained after a single round of panning were analyzed for their binding specificity. The murine VH genes encoding tetanus toxoid-specific Fab fragments were subjected to sequence analysis to identify unique clones and assign VH, DH and JH utilization.

[0110]Many of the protocols described here are standard protocols for the construction of phage display libraries and the panning of phages for binding to an antigen of interest and described in Antibody Phage Display: Methods and Protocols ...

example 3

Silencing of the Mouse Kappa Light Chain Locus

[0125]This example describes the silencing of the mouse endogenous kappa light chain locus. The endogenous kappa locus is modified by homologous recombination in ES cells, followed by the introduction of genetically modified ES cells in mouse embryos to obtain genetically adapted offspring.

[0126]A vector that contains an assembled nucleotide sequence consisting of a part comprising the J-region to 338 by downstream of the J5 gene segment fused to a sequence ending 3′ of the 3′ CK enhancer is used for homologous recombination in ES cells. The assembled sequence is used to delete a genomic DNA fragment spanning from 3′ of the JK region to just 3′ of the 3′ CK enhancer. As a consequence of this procedure, the CK constant gene, the 3′ enhancer and some intergenic regions are removed (see FIGS. 4 and 18).

Construction of the Targeting Vector

[0127]A vector that received 4.5-8 kb flanking arms on the 3′ and 5′ end fused to the deletion segment w...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
volumesaaaaaaaaaa
volumesaaaaaaaaaa
concentrationaaaaaaaaaa
Login to View More

Abstract

Described are transgenic, non-human animals comprising a nucleic acid encoding an immunoglobulin light chain, whereby the immunoglobulin light chain is human, human-like, or humanized. The nucleic acid is provided with a means that renders it resistant to DNA rearrangements and / or somatic hypermutations. In one embodiment, the nucleic acid comprises an expression cassette for the expression of a desired molecule in cells during a certain stage of development in cells developing into mature B cells. Further provided is methods for producing an immunoglobulin from the transgenic, non-human animal.

Description

CROSS-REFERENCE TO RELATED APPLICATION[0001]This application claims the benefit, under 35 U.S.C. §119(e), to U.S. Provisional Patent Application Ser. No. 61 / 133,274, filed Jun. 27, 2008, for “Antibody Producing Non-Human Mammals,” the entire contents of which are hereby incorporated hereby by this reference.TECHNICAL FIELD[0002]The invention relates to the production and use of non-human animals capable of producing antibodies or derivatives thereof, which are expressed from at least partially exogenous nucleic acids (transgenes). Transgenes to produce such transgenic animals and methods to produce such heterologous antibodies; methods and vectors for producing such transgenic animals are disclosed.BACKGROUND OF THE INVENTION[0003]B cells mediate humoral immunity by producing specific antibodies. The basic structural subunit of an antibody (Ab) is an immunoglobulin (Ig) molecule. Ig molecules consist of a complex of two identical heavy (H) and two identical light (L) polypeptide cha...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(United States)
IPC IPC(8): C07K16/18A01K67/00C07K16/00C07K16/10C07K16/22
CPCA01K67/0278C07K16/1282A01K2217/052A01K2217/075A01K2217/15A01K2217/206A01K2227/105A01K2267/01C07K14/47C07K16/248C07K2317/24C12N15/8509A01K67/027C12P21/00A01K2207/15A61P31/14C07K16/005C07K16/10C07K16/22C07K16/2863C07K16/32C07K2317/14C07K2317/31C07K2317/34C07K2317/52C07K2317/55C07K2317/56C07K2317/622C07K2317/76C07K2317/94C07K2319/00A01K67/0275C07K16/462C07K16/00C07K2317/10C07K2317/21C07K2317/51C07K2317/515C07K2317/64G01N33/56966C12N5/10
Inventor HOUTZAGER, ERWINPINTO, RUI DANIELLOGTENBERG, TONTHROSBY, MARK
Owner MERUS NV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com