The present invention provides methods and compositions for generating transgenic animals, including transgenic mammals, as well as plasma cells that allow for cell surface capture of secreted immunoglobulin molecules produced endogenously in the plasma cells.

The present invention describes transgenic animals with human(ized) immunoglobulin loci and transgenes encoding human(ized) Igα and/or Igβ sequences. Of particular interest are animals with transgenic heavy and light chain immunoglobulin loci capable of producing a diversified human(ized) antibody repertoire that have their endogenous production of Ig and/or endogenous Igα and/or Igβ sequences suppressed. Simultaneous expression of human(ized) immunoglobulin and human(ized) Igα and/or Igβ results in normal B-cell development, affinity maturation and efficient expression of human(ized) antibodies.

The invention provides methods and compositions for perturbing gene expression in hematopoietic cell lineages in vivo.

Methods for producing an antibody or an antigen-binding fragment thereof specifically binding to an antigen of interest, methods for inducing proliferation of PBMCs, B cell activation and differentiation, B cell maturation, and/or promoting class switch in an antibody-producing PBMC to produce IgG, compositions for the in vitro immunization and methods for identifying an antibody-enhancing factor for in vitro immunization.

This disclosure relates to growth media and environments for in vitro culturing of cells that produce or are capable of producing antibodies. In certain embodiments, the media comprises IL-6, fibronectin, and typically a saccharide. In certain embodiments, the disclosure contemplates cell culture compositions comprising IL-6 and fibronectin that are derived from proteins secreted from mesenchymal stromal/stem cells (MSCs). In certain embodiments, the disclosure contemplates enclosures comprising culture compositions disclosed herein that are in ambient air or optionally in an environment wherein oxygen is absent or at a low concentration.

In certain aspects, the present invention provides methods for inducing a stable gene modification of a target nucleic acid via homologous recombination in a primary cell, such as a primary blood cell and/or a primary mesenchymal cell. In certain other aspects, the present invention provides methods for enriching a population of genetically modified primary cells having targeted integration at a target nucleic acid. The methods of the present invention rely on the introduction of a DNA nuclease such as a Cas polypeptide and a homologous donor adeno-associated viral (AAV) vector into the primary cell to mediate targeted integration of the target nucleic acid. Also provided herein are methods for preventing or treating a disease in a subject in need thereof by administering to the subject any of the genetically modified primary cells or pharmaceutical compositions described herein to prevent the disease or ameliorate one or more symptoms of the disease.

The invention provides freeze-dried nucleated cells, a method for preparing them, and methods of using them for in vitro assays and in vivo therapeutic treatments. The method for preparing the cells includes incubating cells in the presence of a cryoprotective sugar to load them with the sugar, then lyophilizing them without separating the cells from the cryoprotective sugar. In embodiments, the cells are also loaded with one or more bioactive agents.

Provided herein are methods and systems for bio-printing of three-dimensional organs and organoids. Also provided herein are bio-printed three-dimensional organs and organoids for use in the generation and/or the assessment of immunological products and/or immune responses. Also provided herein are methods and system for bio-printing three-dimensional matrices.

Provided herein are methods for expanding populations of regulatory B cells comprising engineering a population of B cells to express CD40 ligand. Also provided herein are methods of treating immune disorders with the regulatory B cells.

The instant technology generally relates to improved methods for producing antibodies, antibody libraries, hybridomas, hybridoma libraries, etc. For example, these methods increase the number of antigen-specific B cells produced, increase the number of hybridomas, and/or increase the number of monoclonal antibodies that can be made in a given production cycle.