Services

Cell Line Development

Introduction

The CHO-based stable cell lines are designed with GlycoNex proprietary expression vector to achieve time efficient, cost-effective and high yield. Combined with comprehensive analytical platforms, the cell lines are generated with desired post translational modification, productivity and stability as customized. The whole process is under animal component free condition and could be easily scaled up from lab scale to 1000L manufacturing scale.

Monoclonality Verfication

The high producing clones were isolated by cell sorting and the monoclonality was confirmed by well-to-well image scan. With one round cloning, the probability that a cell will be monoclonal is 99.79% at the 95% confidence interval.

Customized N-glycan Profile

The N-glycan profile could not be the result of the clone. In GlycoNex’s platform, we make it as criteria in cell line generation. Combined with upstream and downstream process development, the N-glycan profile is customized for your needs.

Our Commitment

50L Pilot Production

Easy Scale-up – Comparable Cell Growth and Titer Performance


Through the comprehensive process development, GlycoNex provides the scalable and robust processes for scale-up from 1L to 50L pilot production. The cost-effective, efficient, and fully- documented pilot production provides materials for toxicology studies.

 

Antibody Characterization

Introduction

In-depth characterization of therapeutic antibody is important for producing the product with high safety and consistent efficacy. However, due to the inherent heterogeneity of N-glycan and other amino acid modifications, thorough characterization of antibody is a challenging task. With powerful Mass spectrometry (MS) and UPLC techniques developed at GlycoNex, we provide tools for in-depth analyses of mAbs including intact protein, peptide and glycan profiling.

 

Peptide (bottom-up) analysis

We achieve 100% sequence coverage through

 
  • Single protease digestion
  • H+L chain mixture
  • 25 min analytical time (nanoLC-MSMS)
  • High confidence with MS/MS sequencing
  • An optimized 12 hr workflow


 

 

Intact protein analysis

High-accuracy analysis with high resolution mass spectrometer (Mass discrepancy <1 Da for IgG)

 

Glycan profile analysis

 

  • Qualitative analysis by Mass spectrometry
  • Qualitative and quantitative analyses by UPLC and glycan standards


 

 

Functional Evaluation

Activity assessment with in vitro & in vivo Assays
 

Introduction

Functional assays are often employed to obtain quantified information of antibody’s biological activities. Applicable functional assays not only support lead antibody selection but also help characterization of the mechanism of action.

GlycoNex offers multiple types of activity evaluation platforms including ELISA, SPR, flow cytometry, cell-based assays as well as in vivo xenograft models. With a variety of target cell lines in hand, our assay platform helps clients select lead candidates efficiently. Clients are also welcomed to provide their unique cell lines for functional evaluation.

Our experienced scientists and well-designed platforms provide sensitive assays with low noise and high repeatability that produce reliable data with quality management to our clients. GlycoNex is certified according to ISO 9001.

Our Core Competencies

Binding activity

Affinity evaluation
•Ligand binding
Fc receptor binding (FcγRIa, FcγRIIa, FcγRIIIa and FcγRn)
C1q binding

Antibody-Fc receptor interactions activate many cytotoxic pathways, including ADCC (via FcγRIIIa/CD16a), and ADCP (via FcγRIIa/CD32a). C1q is a predominant complement component that engages Fc domain of an antibody and subsequently activates CDC pathway. GlycoNex provides Fcγ Receptors and C1q ELISA binding assays as well as Biacore™ analysis as surrogate methods to evaluate ADCC and CDC activities.


Cell binding assay
•Target cell binding

GlycoNex has multiple cancer cell lines derived from various organs for target screening and functional evaluation. Flow cytometry is applied for cell binding assay, which can serve as a primary screening tool to determine potential target cell lines, and to detect target expression level on cellular surface.


 

Bioactivity

•ADCC
CDC

For therapeutic antibodies, Antibody-Dependent Cell-mediated Cytotoxicity (ADCC) and Complement-Dependent Cytotoxicity (CDC) activities are important indicators to understand their mechanisms and efficacy in target cancer cell lines. With decades of experience in antibody research, GlycoNex is pleased to present to our clients with various well-established platforms to measure ADCC and CDC activities. Parallel FACS analysis can be provided upon request.


•Proliferation assays

GlycoNex provides sensitive colorimetric assays for the determination of the number of viable or dead cells in proliferation and cytotoxicity assays.

Efficacy

•Xenograft models

It is essential to validate the efficacy of drug candidates in vivo. GlycoNex has established various subcutaneous xenograft models. We also offer immunohistochemistry (IHC) analyses to detect the expression of drug targeted antigens and intended markers on multiple tissues.

Overall, these approaches enable our clients to corroborate the anti-tumor activity of developed antibodies in vivo, explore antigen distribution as well as provide guidance in selecting appropriate cancer indications in the future.

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Antibody Engineering

Antibody improvements through 3D modeling and structural engineering
 

Introduction

GlycoNex launches antibody engineering service that incorporates structure-based computational 3D modeling and X-ray crystallography to perform humanization, specificity and affinity improvements. Through the simulated antigen/antibody complex structure, key amino acid residues that are involved in Ag-Ab interaction or important for structural stability are resolved. By mutational energy calculations, specific amino acid switch at one or more of these residues or nearby positions which leads to improvement of the antibody will be resolved. This platform technology not only reduces the potential risk of immunogenicity but also optimizes antibody specificity and affinity. Customers can also integrate our antibody production services to obtain engineered antibody for further in vitro and in vivo functional assays.