Monoclonal antibodies (mAb)

Monoclonal antibodies are identical immunologically active proteins which specifically bind to the same epitope. They are monoclonal as they arise from a single B-lymphocyte cell line (cell clone). Polyclonal antibodies are secreted by different B cell lineages. These bind to multiple epitopes and are therefore less specific.

When fighting diseases, it is this antigen-specific trait that is necessary to achieve effective treatment of diseases with controlled risk of adverse effects. Specific molecules (antigens) are first identified that have a crucial role in the course of a disease. These are often cell membrane receptors or transport and signal proteins. These then serve as the epitope for the monoclonal antibodies to bind to. The hybridoma technology, for which César Milstein and Georges Köhler received the Nobel Prize in 1984, is used to produce these monoclonal antibodies.

Monoclonal antibodies HPLC columns

In order to adequately examine biological samples, a qualitative separation of the components is necessary. Basic techniques like chromatography are used. High resolution and reproducibility are the most important requirements for the chromatographic separation of biomolecules.

Thermo Scientific MAbPac Columns

Thermo Fisher Scientific offers a series of HPLC columns for the processing of biological samples. In addition to protein and peptide HPLC columns, monoclonal antibody HPLC columns are also offered. They enable the characterisation and quantification of monoclonal antibodies, titer analysis and the separation of antibody fragments, aggregates and related biologicals. They also serve to monitor the stability and to control the efficiency and safety of antibodies.

Thermo Scientific distributes five different product lines of its MAbPac LC columns:

  • Reversed phase monoclonal antibody columns
  • Use reversed-phase conditions for higher resolution separation of intact mAbs and fragments by HPLC and LC-MS.
  • Products: MAbPac RP LC columns
  • Hydrophobic interaction chromatography (HIC) mAb columns
  • Use these columns for mAb, mAb fragment, antibody-drug aggregate and related biologic separations.
  • Products: MAbPac HIC LC columns
  • Affinity monoclonal antibody columns
  • Use these columns for fast and accurate titer analysis of monoclonal antibodies in harvest cell cultures.
  • Products: MAbPac Protein A LC columns
  • Strong cation exchange monoclonal antibody columns
  • Use these columns for high-resolution, high-efficiency analysis of mAbs and associated variants.
  • Products: MAbPac SCX LC columns
  • Size exclusion monoclonal antibody columns
  • Choose these columns for optimal mAb analysis, including the high-resolution separation of monomers, aggregates and fragments.
  • Products: MAbPac SEC LC columns

The following table should help you find the right MAbPac LC column for your mAb analysis. Please contact us, if you have any questions..



Columns & Buffers

mAb Capture & titer analysis

  • mAb titer determination (concentration) & screening
  • mAb capture for analysis workflows
Aggregate analysis
  • Routing screening for mAb aggregates and fragments

Charge variant analysis

  • Routine charge variant profiling/screening, including lysine truncation, deamidation & acylation

Methionine & tryptophan oxidation

  • Targeted analysis of methionine and tryptophan oxidation

Antibody drug conjugate (ADC) analysis

  • ADC DAR analysis

Intact mAb & fragment analysis

  • Light chain (LC) and heavy chain (HC) analysis
  • Fab and Fc analysis
  • scFc and F(ab’)2 analysis

Sequence & structural analysis

  • Primary sequence analysis<
  • peptide mapping
  • peptide & glycopeptides structural & linkage analysis

Glycan profiling

  • Profiling of released glycans

Hybridoma technology

Hybrodoma technology produces monoclonal antibodies by fusing antibody-producing B-lymphocytes with myleoma cells to combine properties of both cell types. First, mice are injected with an antigen specially selected for the targeted monoclonal antibodies (receptor or signal proteins) to bind to. As an immune response B-lymphocytes develop in the mouse and produce antibodies that bind to the injected antigen. The B lymphocytes are isolated from the spleen and fused with plasma cells from a myeloma which are immortal tumor cells as they can multiply extremely quickly. Hybridomas are formed which combines the longevity and reproductivity of the myeloma with the antibody-producing ability of the B lymphocytes. This creates highly stable, long-lasting cells that continuously produce monoclonal antibodies.

Medical use

The diverse use of monoclonal antibodies in diagnostics and therapy has made them increasingly indispensable since their first discoveries in the 1970s.

In the laboratory, they are used for immunohistological procedures or ELISA and enable better reproducibility than polyclonal antibodies. They are also used to identify various tumors in in-vivo diagnostic procedures.

They are also increasingly used in the therapy of diseases in which key molecules have been identified against which monoclonal antibodies develop.

These therapeutically used antibodies are generally named according to their origin: In murine antibodies, both the constant and the variable region consist of mouse proteins. These antibodies could only be used to a very small extent as they were recognised and fought against by the human immune system as antigenic structures. The development to fully human antibodies via chimeric antibodies (contain only about 30 % murine proteins) and humanised antibodies (90 % from human proteins) nowadays enables promising use in many different indications without an increased risk of immune system defense reactions.

Examples include infliximab as an antibody against TNF-alpha in psoriasis, rituximab as an antibody against CD20 for the treatment of non-Hodgkin lymphoma or omalizumab, which is used in severe bronchial asthma and binds to IgE to reduce anaphylactic reactions.