Identification of HBsAg-specific antibodies from a mammalian cell displayed full-length human antibody library of healthy immunized donor

Li, Changzheng; Liang, Zhongkun; Chen, Zhen-Rui; Lou, Haibo; Zhou, Yizhang; Zhang, Zhehuan; Yu, Fei; Liu, Shuwen; Zhou, Yuanping; Wu, Shuai; Wen-ling, Zheng; Tan, Wanlong; Jiang, Shibo; Zhou, Chen

doi:10.1038/cmi.2011.55

Cited by 15 publications

(10 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Site-specific integration of transgenes directed by Flp recombinase using the commercial "Flp-In" system has previously been described. 10,11 In direct comparison with the nuclease-directed system presented here, we found the Flp-In system to be deficient (see supplementary section), mirroring the rather limited success both in the original publications and subsequently by others. 12 This is likely due to the fact that the Flp-In system is designed for accurate integration in a limited number of clones rather than large library construction.…”

Section: Introductionsupporting

confidence: 59%

A comprehensive search of functional sequence space using large mammalian display libraries created by gene editing

Parthiban¹,

Perera²,

Sattar³

et al. 2019

mAbs

View full text Add to dashboard Cite

The construction of large libraries in mammalian cells allows the direct screening of millions of molecular variants for binding properties in a cell type relevant for screening or production. We have created mammalian cell libraries of up to 10 million clones displaying a repertoire of IgG-formatted antibodies on the cell surface. TALE nucleases or CRISPR/Cas9 were used to direct the integration of the antibody genes into a single genomic locus, thereby rapidly achieving stable expression and transcriptional normalization. The utility of the system is illustrated by the affinity maturation of a PD-1-blocking antibody through the systematic mutation and functional survey of 4-mer variants within a 16 amino acid paratope region. Mutating VH CDR3 only, we identified a dominant "solution" involving substitution of a central tyrosine to histidine. This appears to be a local affinity maximum, and this variant was surpassed by a lysine substitution when light chain variants were introduced. We achieve this comprehensive and quantitative interrogation of sequence space by combining high-throughput oligonucleotide synthesis with mammalian display and flow cytometry operating at the multi-million scale.

show abstract

Section: Introductionsupporting

confidence: 59%

A comprehensive search of functional sequence space using large mammalian display libraries created by gene editing

Parthiban¹,

Perera²,

Sattar³

et al. 2019

mAbs

View full text Add to dashboard Cite

show abstract

“…As shown in Figure 5, the induction of anti-HBV specific immune response in the liverdraining LNs was observed, and the adoptive transfer of cells from the liver-draining LN of HBV plasmid-injected mice into Rag1 2/2 mice can effectively decrease serum HBsAg levels in recipient mice. Although a previous study indicated that production of HBsAg-specific antibodies is important in the management of HBV infection, 25 whether the anti-HBs are produced in the liver-draining LNs needs further study. Collectively, we characterized the involvement of the liverdraining LNs in the immune response to HBV and recognized them as contributing factors to HBV elimination.…”

Section: Induction Of Anti-hbv Responses In the Liver-draining Lnsmentioning

confidence: 99%

Characterization of the liver-draining lymph nodes in mice and their role in mounting regional immunity to HBV

Zheng

Tian

2013

Cell Mol Immunol

View full text Add to dashboard Cite

The lymphatic system is important in mounting an immune response to foreign antigens and tumors in humans and animal models. The liver produces a large amount of lymph, and its lymphatic system is divided into three major components: the portal, sublobular and superficial lymphatic vessels. Despite the fact that mice are the most commonly used laboratory animals, detailed descriptions of the anatomical location and function of the lymph nodes (LNs) that drain the liver are surprisingly absent. In this study, we found that the portal and celiac LNs adjacent to mouse liver were stained with Evans blue within 5-8 min. Enhanced green fluorescence protein (EGFP)-positive cells from the liver also drained into the two aforementioned LNs. These data indicate that the portal and celiac LNs drain the mouse liver. Lymphadenectomy of the identified liver-draining LNs resulted in hepatitis B virus (HBV) persistence in immunocompetent mice compared with the sham group. In addition, the frequencies of CD81 T cells and dendritic cells (DCs) increased significantly in the liver-draining LNs after hydrodynamic injection of HBV plasmid. Liver-draining LN cells in HBV plasmid-injected mice also showed significant antigen-specific proliferation in response to stimulation with recombinant hepatitis B core antigen in vitro. Adoptive transfer of these cells into Rag1 2/2 mice induced a reduction in the serum concentration of hepatitis B surface antigen (HBsAg) compared to liver-draining LN cells in uninjected mice. Altogether our data characterize the liver-draining LNs and provide evidence that the liver-draining LNs induce an anti-HBV-specific immune response responsible for HBV clearance.

show abstract

“…As the need for therapeutic antibodies for treatment of human diseases increases, platforms will transition more towards mammalian cell display, requiring corresponding advancement in all aspects of library construction. We reported the development of a unique mammalian display platform [13] - [18] . In our platform, a trans-membrane (TM) domain was fused to the 3′end of heavy chain in frame to display full-length antibody on mammalian cell surface.…”

Section: Discussionmentioning

confidence: 99%

“…Dual expression vectors containing both heavy and light chain genes can be constructed in single four-way ligation [14] . Several full-length fully human antibody display libraries with a combinational diversity of 10 9 have been constructed [15] – [17] , and antigen-specific antibodies have been identified from one of the constructed libraries [18] .…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Expression of Displayed and Secreted Antibodies for Antibody Screen

et al. 2013

Self Cite

View full text Add to dashboard Cite

The display of full-length antibody on the cell surface was achieved by fusing a transmembrane domain of the platelet-derived growth factor receptor (PDGFR) to the C-terminus of the heavy chain constant region. We also incorporated a furin cleavage site between the constant region and PDGFR transmembrane domain to obtain secreted antibodies. As a result, antibodies can be expressed simultaneously on the cell surface in a membrane-anchored version for screening and selecting through fluorescence-activated cell sorting (FACS) analysis, as well as in conditioned medium in a secreted version for function analysis.

show abstract

Identification of HBsAg-specific antibodies from a mammalian cell displayed full-length human antibody library of healthy immunized donor

Cited by 15 publications

References 22 publications

A comprehensive search of functional sequence space using large mammalian display libraries created by gene editing

A comprehensive search of functional sequence space using large mammalian display libraries created by gene editing

Characterization of the liver-draining lymph nodes in mice and their role in mounting regional immunity to HBV

Simultaneous Expression of Displayed and Secreted Antibodies for Antibody Screen

Contact Info

Product

Resources

About