HW #9c Affinity Chromatography

Which of the following are lectins that can be used in affinity chromatogrpahy to bind to sugar containing molecules?

1. Concanacalin A 2. Wheat germ agglutinin

Match the biochemical pairs of materials that could be used in affinity chromatography 1. Protein (such as a histone) 2. Antigen 3. Enzyme 4. Receptor

1. Nucleic acid 2. Antibody 3. Substrate 4. Ligand

Match the following specific combination of affinity interactions commonly used in this technique 1. Lectin 2. Poly-A 3. Metal ions 4. Proteins A and G 5. Avidin 6. Enzymes

1. Polysaccharide 2. RNA containing poly (U) sequence 3. Recombinant fusion proteins with His-tag 4. Immunoglobulins 5. Biotin 6.Substrate analogue

(T/F) Affinity chromatogrpahy is a method of separating biochemical mixture based on a hightly speciic interaction between antigen and antibody, enzyme and substrate, receptor and ligand, or protein and nucleic acid. It is a type of chromatographic laboratory technique used for purifying biological moleucles within a mixture by exploiting molecular properites.

True

(T/F) Possibly the most common use of affinity chromatography is for the purification of recombinant proteins.

True

(T/F)Alkaline phosphatase from E. coli can be purified using a DEAE-Cellulose matrix. A. phosphatase has a slight negative charge, allowing it to weakly bind to the positively charged amine groups in the matrix. The enzyme can then be eluted out by adding buffer with higher salt concentrations.

True

(T/F)Immunoaffinity Chromatography is the affinity purification of antibodies from blood serum which is known to contain antibodies against a specific antigen

True

(T/F)Most monoclonal antibodies have been purified using affinity chromatography based on immunoglobulin-specific Protein A or Protein G, derived from bacteria

True

(T/F)The high selectivity of affinity chromatogrpahy is caused by allowing the desired molecule to interact with the stationary phase and be bound within the column in order to be separated from the undesired material which will not interact and elute first.

True

(T/F)The starting material for separation using affinity chromatography can be a crude, heterogeneous group of molecuels in a whole cell extract, such as a cell lysate, growth medium or blood serum

True