Affinity Chromatography: Test Your Knowledge with Important MCQs
Which of the following techniques is primarily based on specific molecular interactions for separation? a) Size exclusion chromatography b) Affinity chromatography c) Ion exchange chromatography d) Hydrophobic interaction chromatography
Solution: b) Affinity chromatography Reason: Affinity chromatography relies on specific molecular interactions, such as binding between a ligand and its target molecule, for separation.
What is the purpose of the ligand in affinity chromatography? a) To increase column capacity b) To decrease resolution c) To provide stability to the column d) To specifically bind the target molecule
Solution: d) To specifically bind the target molecule Reason: The ligand in affinity chromatography is chosen to specifically bind the target molecule of interest, enabling its isolation from other components in the mixture.
Which of the following statements about affinity chromatography is true? a) It separates molecules based on their size b) It separates molecules based on their charge c) It separates molecules based on specific interactions with a ligand d) It separates molecules based on their hydrophobicity
Solution: c) It separates molecules based on specific interactions with a ligand Reason: Affinity chromatography utilizes specific interactions between the target molecule and the immobilized ligand for separation.
In affinity chromatography, what role does the elution buffer play? a) To increase binding affinity b) To decrease specificity c) To disrupt the binding interactions between the ligand and the target molecule d) To increase column capacity
Solution: c) To disrupt the binding interactions between the ligand and the target molecule Reason: The elution buffer is used to disrupt the specific interactions between the ligand and the target molecule, facilitating the release of the target molecule from the column.
Which of the following is a common application of affinity chromatography? a) Separating proteins based on size b) Separating proteins based on charge c) Isolating specific antibodies d) Separating proteins based on hydrophobicity
Solution: c) Isolating specific antibodies Reason: Affinity chromatography is commonly used to isolate specific proteins or molecules, such as antibodies, based on their specific binding interactions with the immobilized ligand.
What is typically used to immobilize the ligand in affinity chromatography? a) A gel matrix b) A metal ion c) The chromatography matrix d) A protein A/G resin
Solution: c) The chromatography matrix Reason: The ligand is often immobilized onto the chromatography matrix, allowing for specific interactions with the target molecule as it flows through the column.
Which of the following factors can influence the specificity of binding in affinity chromatography? a) Temperature b) pH c) Ionic strength d) All of the above
Solution: d) All of the above Reason: Factors such as temperature, pH, and ionic strength can all influence the specificity of binding between the ligand and the target molecule in affinity chromatography.
Affinity chromatography is particularly useful for isolating molecules with: a) Low specificity b) High purity c) Low molecular weight d) High solubility
Solution: b) High purity Reason: Affinity chromatography enables the isolation of molecules with high purity by selectively binding the target molecule and minimizing contamination from other components.
Which of the following is commonly used as a ligand in affinity chromatography for purifying His-tagged proteins? a) Streptavidin b) Biotin c) Protein A d) Ni-NTA
Solution: d) Ni-NTA Reason: Ni-NTA (nickel-nitrilotriacetic acid) is commonly used as a ligand in affinity chromatography for purifying proteins containing a histidine (His) tag.
What is the primary driving force behind the separation in affinity chromatography? a) Electrostatic interactions b) Hydrophobic interactions c) Van der Waals forces d) Specific binding interactions
Solution: d) Specific binding interactions Reason: The specific binding interactions between the ligand and the target molecule are the primary driving force behind separation in affinity chromatography.
Which of the following is a limitation of affinity chromatography? a) Low specificity b) High cost of ligands c) Limited capacity d) Non-specific binding
Solution: c) Limited capacity Reason: Affinity chromatography may have a limited capacity for binding target molecules, especially in large-scale purifications.
What role does the spacer arm play in affinity chromatography? a) It stabilizes the ligand b) It increases the capacity of the column c) It prevents steric hindrance d) It enhances the specificity of binding
Solution: c) It prevents steric hindrance Reason: The spacer arm in affinity chromatography helps prevent steric hindrance between the ligand and the target molecule, allowing for efficient binding.
Which of the following is a common ligand used for affinity chromatography of nucleic acids? a) Streptavidin b) Ni-NTA c) Protein A/G d) Biotin
Solution: d) Biotin Reason: Biotin is frequently used as a ligand for affinity chromatography of nucleic acids due to its high affinity for streptavidin, which can be immobilized on the chromatography matrix.
Which technique is often used to elute the target molecule from the affinity column? a) Increasing temperature b) Changing pH c) Adding salt d) All of the above
Solution: d) All of the above Reason: Different elution conditions such as changing pH, adding salt, or increasing temperature can disrupt the binding interactions and facilitate elution from the column.
Which of the following is a characteristic feature of affinity chromatography? a) Rapid separation b) Non-specific interactions c) High specificity d) Low resolution
Solution: c) High specificity Reason: Affinity chromatography offers high specificity, enabling the isolation of specific molecules from complex mixtures with minimal contamination.
Which type of chromatography primarily relies on the specific interaction between a target molecule and an immobilized ligand? a) Gas chromatography b) Thin-layer chromatography c) Affinity chromatography d) Reverse-phase chromatography
Solution: c) Affinity chromatography Reason: Affinity chromatography is unique in its reliance on specific interactions between a target molecule and an immobilized ligand for separation.
What is the primary purpose of washing the column in affinity chromatography after sample application? a) To increase column capacity b) To decrease specificity c) To remove nonspecifically bound molecules d) To enhance elution efficiency
Solution: c) To remove nonspecifically bound molecules Reason: Washing the column helps remove molecules that are not specifically bound to the immobilized ligand, thereby improving the purity of the eluted sample.
Affinity chromatography is particularly suitable for isolating proteins with: a) Low purity b) Low molecular weight c) High specificity d) Low solubility
Solution: c) High specificity Reason: Affinity chromatography excels in isolating proteins with high specificity, allowing for the purification of target proteins from complex mixtures.
Which of the following statements about affinity chromatography is true? a) It is based on nonspecific interactions b) It separates molecules based on their size c) It relies on specific binding interactions between molecules d) It is suitable for isolating proteins solely based on their size
Solution: c) It relies on specific binding interactions between molecules Reason: Affinity chromatography separates molecules based on specific binding interactions between the ligand and the target molecule, ensuring high specificity.
What is typically used as the stationary phase in affinity chromatography? a) A mobile liquid phase b) A gel matrix c) A gas phase d) The chromatography matrix
Solution: d) The chromatography matrix Reason: The chromatography matrix serves as the stationary phase in affinity chromatography, providing a solid support for immobilizing the ligand.
Which of the following is an advantage of affinity chromatography? a) Low resolution b) Non-specific binding c) High purity d) Rapid separation
Solution: c) High purity Reason: Affinity chromatography offers high purity in isolating target molecules, minimizing contamination from other components present in the sample.
Affinity chromatography is commonly used in which stage of protein purification? a) Primary purification b) Secondary purification c) Tertiary purification d) Quaternary purification
Solution: a) Primary purification Reason: Affinity chromatography is often used as the initial step in protein purification to isolate the target protein from complex mixtures.
Which of the following techniques is NOT a type of affinity chromatography? a) Immobilized metal ion affinity chromatography (IMAC) b) Affinity precipitation chromatography c) Hydrophobic interaction chromatography (HIC) d) Lectin affinity chromatography
Solution: c) Hydrophobic interaction chromatography (HIC) Reason: Hydrophobic interaction chromatography separates molecules based on their hydrophobicity, not specific binding interactions.
What is the primary role of the ligand in affinity chromatography? a) To provide stability to the column b) To increase column capacity c) To specifically bind the target molecule d) To enhance elution efficiency
Solution: c) To specifically bind the target molecule Reason: The ligand in affinity chromatography is chosen for its ability to selectively bind the target molecule of interest.
Which technique is commonly used to detect the eluted proteins in affinity chromatography? a) Gel electrophoresis b) Western blotting c) UV-visible spectroscopy d) Mass spectrometry
Solution: c) UV-visible spectroscopy Reason: UV-visible spectroscopy is often used to detect proteins eluted from affinity chromatography columns based on their absorbance properties.
What is the primary driving force for elution in affinity chromatography? a) Increasing specificity b) Decreasing column capacity c) Disrupting specific binding interactions d) Enhancing column stability
Solution: c) Disrupting specific binding interactions Reason: Elution in affinity chromatography is achieved by disrupting the specific binding interactions between the ligand and the target molecule.
Affinity chromatography is particularly effective for purifying proteins with: a) Low specificity b) High purity c) Low molecular weight d) Low solubility
Solution: b) High purity Reason: Affinity chromatography allows for the isolation of proteins with high purity by selectively binding the target molecule and minimizing contamination from other components.
What role does the spacer arm play in affinity chromatography? a) Enhancing column capacity b) Increasing elution efficiency c) Preventing steric hindrance d) Improving column stability
Solution: c) Preventing steric hindrance Reason: The spacer arm in affinity chromatography helps prevent steric hindrance between the ligand and the target molecule, facilitating efficient binding.
Which of the following statements regarding elution in affinity chromatography is correct? a) Elution is achieved by increasing the size of the column b) Elution is achieved by decreasing the pH of the elution buffer c) Elution is achieved by disrupting specific binding interactions d) Elution is achieved by enhancing column stability
Solution: c) Elution is achieved by disrupting specific binding interactions Reason: Elution in affinity chromatography involves disrupting the specific binding interactions between the ligand and the target molecule to release the bound molecule from the column.
Which of the following is NOT a common application of affinity chromatography? a) Purification of enzymes b) Isolation of specific antibodies c) Separation of proteins based on size d) Capture of specific proteins from complex mixtures
Solution: c) Separation of proteins based on size Reason: Affinity chromatography is not primarily used for separating proteins based on size but rather for isolating specific molecules based on their affinity for a particular ligand.
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