Study Guide #4

Monoclonal antibodies

Proteins made in a laboratory that act like antibodies in the body.

Hybridoma

Method for producing large numbers of identical antibodies.

ELISA

Test to detect and quantify antibodies in the blood.

Flow cytometer (FCM)

Technology that provides rapid multi-parametric analysis of single cells in solution.

Fluorescence-activated cell sorter (FACS)

Specialized type of flow cytometry.

Affinity

The strength of the interaction between the antigen-binding site on an antibody and the epitope on an antigen.

Avidity

Total strength required for the interaction between a multivalent antibody and multiple antigenic epitopes.

Agglutination

A reaction in which particles suspended in a liquid collect into clumps, especially as a serological response to a specific antibody.

Biotin

Promoted for hair, skin, and nail health.

Myeloma cells

Abnormal plasma cells that build up in the bone marrow and form tumors in many bones of the body.

PAGE

High-resolution technique commonly used for separating a mixture of proteins according to their molecular weight.

Polyclonal antibody

A complex mixture of several antibodies produced by different B-cell clones of an animal.

Sodium dodecyl sulfate

Detergent agent that breaks disulphide bonds of proteins, disrupting their tertiary structure.

Streptavidin

Tetrameric protein commonly used in biotechnology as a stable linker due to its strong affinity for biotin molecules.

Western blotting

Laboratory technique used to detect a specific protein in blood or tissue.

Biotin interference

Elevated biotin concentrations can falsely increase analyte concentration in competitive immunoassays.

Animal antisera limitations

Limited supply and ethical concerns regarding euthanizing animals for antiserum.

Monoclonal antibodies advantages

Produced in large amounts, less expensive.

Pooled human Ig advantages

Ensures diversity of antibodies.

Flow cytometry components

Fluidics, optics, and electronics.

Forward scatter

Measures scatter along the path of the laser.

Side scatter

Measures scatter at a ninety-degree angle.

Absorbance spectra

Atoms absorb energy.

Emission spectra

Radiation emitted by excited atoms.

Common fluorochromes

VioBlue (Purple), FITC (Blue/Green), APC (Red/Orange).

Dual labeling

Concept involving simultaneous detection of two different markers.

Lateral flow technology

Used in home pregnancy tests to measure human chorionic gonadotropin hormone.

ELISA formats

Direct, indirect, and sandwich formats designed for quantitative measurement.

Sodium dodecyl sulfate in Western blotting

Imparts a relative negative charge to proteins for size separation.

Polyacrylamide gel electrophoresis

Technique for separating proteins based on size.