RenLite

RenLite™ (Common Light Chain) Mouse: Empower Bispecific Antibody Development

Schematic of Humanization in The RenLite™ Mice

The RenLite™ mouse is a genetically-engineered model with complete humanization in the variable region of the heavy chains, while maintaining a fully-humanized common light chain strategically engineered into the antibody gene. 

Whole mouse heavy chain gene was replaced with full human heavy chain gene in RenLite™ mice.

Whole mouse light chain gene was replaced with single human VK and VJ gene in RenLite™ mice.

Heavy Chain Germline Usage

Heavy chain IGHV germline usage of naive RenLite™ mice

Heavy chain IGHV germline usage of naive RenLite™ mice

Heavy chain IGHD germline usage of naive RenLite™ mice

Heavy chain IGHJ germline usage of naive RenLite™ mice

Human-like Antibody Repertoire

Heavy chain CDR3 length distribution from naive Renite™ mice

Result

The heavy chain CDR3 length distribution was determined by NGS sequencing. As shown in this figure, the median length of heavy chain CDR3 is 14aa, which is consistent with the median length of human heavy chain CDR3.

Heavy chain CDR3 amino acid usage frequency of naive RenLite™ mice

Result

Using 17aa length as an example, the amino acid composition of heavy chain CDR3 was analyzed. Multiple patterns were observed, including the increasing frequency of tyrosine usage.

Heavy chain CDR3 amino acid usage frequency of naive RenLite™ mice

Result

Increasing usage of DH2 germline family which contains Cysteine as heavy chain CDR3 gets longer, resembling a human heavy chain CDR3.

Highly Similar Immune Responses

Result

After three immunizations with antigen A and B, sera titer of RenLite™ and wild type mice were analyzed by ELISA. The results showed that the immune response of RenLite™ and wild type mice were similar.

Result

After three immunizations with antigen A and B, sera titer of RenLite™ and wild type mice were analyzed by ELISA. The results showed that the immune response of RenLite™ and wild type mice were similar.

    Validation Data

Body & Spleen Weight Measurement


Result

Body weight and spleen weight are comparable between RenLite™ and wild type mice.


Result

Body weight and spleen weight are comparable between RenLite™ and wild type mice.

Immune Cell Profiling

Comparison of immune cell population in spleen between RenLite™ and wild type mice.

Result

The percentage of B cells in the spleen of RenLite™ mice is slightly lower than wild type mice. This is mainly due to the limited light chain choice during the B cell maturation. When the heavy chain does not pair with fixed light chain efficiently, the B cells do not mature properly.

B Cell Development Comparison

IgM and IgD expression on B220+ splenocytes were analyzed by flow cytometry. Transitional type 1 (T1, B220+IgM+IgD), Transitional type 2 (T2, B220+IgM+IgD+), Mature (M, B220+IgMlowIgD+) B cell population were quantitatively analyzed.

Result

No significant difference was observed between RenLite™ mice and wild type mice.

CD21 and CD23 expression on B220+ splenocytes were analyzed by flow cytometry. Marginal-zone (MZ, B220+CD21+CD23) and Follicular (FO, B220+CD21lowCD23+) B cells were quantitatively analyzed.

Result

No significant difference was observed between RenLite™ mice and wild type mice.

B Cell Development in Bone Marrow Analysis

B cell precursor subpopulations in bone marrow were analyzed by flow cytometry using B220, CD43 and IgM expression. Percentage of Pro-B-cell (B220lowCD43highIgMlow ), Pre-B-cell (B220lowCD43intIgMlow), and Immature-B-cell (B220highCD43lowIgMhigh) in live cells were shown.

Result

No significant difference was observed between RenLite™ and wild type mice in all 3 subpopulations.

Serum immunoglobulin Isotope & IgG Subtype Analysis

Serum concentrations of IgA, IgM and IgG subtypes of RenLite™ and wild type mice were measured by ELISA. Sera were equally diluted between two groups of mice.

Result

The serum levels of IgG2b, IgG2c and IgG3 of RenLite™ mice were lower than wild type mice. Serum levels of IgA, IgM and IgG1 of RenLite™ mice were either at the same level or slightly higher than those of wild type mice.

Serum concentrations of IgA, IgM and IgG subtypes of RenLite™ and wild type mice were measured by ELISA. Sera were equally diluted between two groups of mice.

Result

The serum levels of IgG2b, IgG2c and IgG3 of RenLite™ mice were lower than wild type mice. Serum levels of IgA, IgM and IgG1 of RenLite™ mice were either at the same level or slightly higher than those of wild type mice.

Serum concentrations of IgA, IgM and IgG subtypes of RenLite™ and wild type mice were measured by ELISA. Sera were equally diluted between two groups of mice.

Result

The serum levels of IgG2b, IgG2c and IgG3 of RenLite™ mice were lower than wild type mice. Serum levels of IgA, IgM and IgG1 of RenLite™ mice were either at the same level or slightly higher than those of wild type mice.

Serum concentrations of IgA, IgM and IgG subtypes of RenLite™ and wild type mice were measured by ELISA. Sera were equally diluted between two groups of mice.

Result

The serum levels of IgG2b, IgG2c and IgG3 of RenLite™ mice were lower than wild type mice. Serum levels of IgA, IgM and IgG1 of RenLite™ mice were either at the same level or slightly higher than those of wild type mice.

Serum concentrations of IgA, IgM and IgG subtypes of RenLite™ and wild type mice were measured by ELISA. Sera were equally diluted between two groups of mice.

Result

The serum levels of IgG2b, IgG2c and IgG3 of RenLite™ mice were lower than wild type mice. Serum levels of IgA, IgM and IgG1 of RenLite™ mice were either at the same level or slightly higher than those of wild type mice.

Serum concentrations of IgA, IgM and IgG subtypes of RenLite™ and wild type mice were measured by ELISA. Sera were equally diluted between two groups of mice.

Result

The serum levels of IgG2b, IgG2c and IgG3 of RenLite™ mice were lower than wild type mice. Serum levels of IgA, IgM and IgG1 of RenLite™ mice were either at the same level or slightly higher than those of wild type mice.