The correct answer is (1) iv. Affinity chromatography.
Introduction
Enzyme purification is a multistep process designed to isolate proteins from complex biological mixtures. Each stage serves to increase the purity and specific activity of the desired enzyme, but not all steps contribute equally. Effective purification is typically identified by dramatic increases in specific activity and reduction of contaminating proteins. This guide analyzes a typical purification table to identify the most effective purification step—in this case, affinity chromatography.
Purification Table Analysis
| Step | Protein (mg) | Activity (units) |
|---|---|---|
| Crude Extract | 20,000 | 4,000,000 |
| Salt precipitation | 5,000 | 3,000,000 |
| Ion-exchange | 200 | 800,000 |
| Affinity chromatography | 50 | 750,000 |
| Size-exclusion chromatography | 45 | 675,000 |
The effectiveness of a step is measured by greatest increase in specific activity:
Specific activity=Activity (units)Protein (mg)
Specific Activity Change
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Before affinity chromatography: Ion-exchange (III): 800,000/200=4,000 units/mg
-
After affinity chromatography (IV): 750,000/50=15,000 units/mg
Affinity step boosts specific activity almost 4-fold, reflecting comprehensive removal of contaminants and retention of most enzyme activity.
Why Affinity Chromatography Is Best
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Design: Uses specific ligand binding to isolate only the enzyme.
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Effectiveness: Yields highest purity and specific activity over other methods, evident from large protein reduction with minimal activity loss.
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Result: Maximally enriches enzyme, removes non-specific proteins and inhibitors.
Explanation of Other Steps
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Salt precipitation and ion-exchange remove some contaminants but less selectively; specific activity rises more modestly.
-
Size-exclusion chromatography gives only slight improvement in purity.
Summary Table for Step Effectiveness
| Step | Specific Activity (units/mg) |
|---|---|
| Crude Extract | 200 |
| Salt precipitation | 600 |
| Ion-exchange | 4,000 |
| Affinity chromatography | 15,000 |
| Size-exclusion chromatography | 15,000 |
Conclusion
Affinity chromatography (step iv) stands as the most effective purification technique in this process due to its sharp increase in specific activity with minimal activity loss. Its molecular selectivity ensures superior enzyme enrichment compared to all other steps.
44 Comments
yashika
September 12, 2025Affinity chromatogeaphy
Mahima Sharma
September 13, 2025Affinity chromatography
Rishita
September 12, 2025Affinity chromatography is the right answer
Vijay Marmat
September 12, 2025Affinity chromatography ✔️
Kirti Agarwal
September 12, 2025Affinity chromatography is best technique for enzyme purification
Khushi Vaishnav
September 12, 2025The most effective purification technique is affinity chromatography
Khushi Vaishnav
September 12, 2025The most effective purification technique is affinity chromatography .
Sakshi yadav
September 12, 2025Affinity chromatography
anjani sharma
September 12, 2025Affinity chromatography is the most effective technique
Kajal
September 12, 2025Affinity chromatography
Neelam Sharma
September 12, 2025Affinity chromatography for enzyme purification kyunki esk ley speci activity jyada h so enzyme purification k ley most effective tec. Is affinity chromatography
Mitali saini
September 13, 2025The correct answer is (1) iv. Affinity chromatography
Varsha Tatla
September 13, 2025Affinity chromatography
Anju
September 13, 2025Ans: affinity chromatography because it separate the molecule with high affinity
Savita Garwa
September 13, 2025Affinity chromatography is best technique for enzyme purification
Kanica Sunwalka
September 13, 2025specific activity = units /mg
on calculation , affinity chromatography have high specific activity ( with minimal activity loss )
therefore its the most effective purification technique .
Kajal
September 13, 2025Option A is correct as affinity chromatography is best for enzyme purification due to its design, effectiveness and result…
Konika Naval
September 13, 2025Affinity chromatography
Aartii sharma
September 13, 2025Affinity chromatography
Asha Gurzzar
September 13, 2025Affinity chromatography
Aakansha sharma Sharma
September 13, 2025specific activity =activity (units) / protein (mg)
on calculation , affinity chromatography have high specific activity ( with minimal activity loss ) So it is most effective purification technique .
MOHIT AKHAND
September 13, 2025(1) Affinity chromatography
Parul Yadav
September 14, 2025The most effective purification step is affinity chromatography as it has increased specific activity by
4-fold.
Deekshita
September 14, 2025Affinity chromatography
Pratibha Jain
September 14, 2025Affinity chromatography is best technique for enzyme purification
Payal Gaur
September 14, 2025Affinity chromatography for enzyme purification
Meera gurjar
September 14, 2025Affinity
Meera gurjar
September 14, 2025By Affinity
Pallavi Ghangas
September 14, 2025Affinity chromatography is best out of all as it gives high yield and is specific and remove non specific protein
anjani sharma
September 14, 2025Affinity chromatography is the best technique as it removes non-specific proteins and inhibitors
Palak Sharma
September 14, 2025specific activity for affinity chromatography is the highest. So it is the best technique.
Dharmpal Swami
September 14, 2025Affinity chromatography
Lokesh Kumawat
September 14, 2025Affinity chromatography
Priya dhakad
September 14, 2025Affinity chromatography is the most effective purification technique.
Divya rani
September 14, 2025Affinity chromatography is the best technique for purification of enzyme or protein. It maintain the specific Binding and dissociate the non specific Binding.
Vanshika Sharma
September 15, 2025Affinity chromatography
Bhawna Choudhary
September 15, 2025Affinity chromatography
Sheikh Aleen
September 15, 2025Affinity chromatography
Soniya Shekhawat
September 15, 2025Affinity chromatography is best technique for enzyme purification.
Nilofar Khan
September 16, 2025correct answer is 1
Affinity chromatography is best tech. For enzyme purification
Mohd juber Ali
September 16, 2025Option 1
Affinity chromatography
Muskan Yadav
September 17, 2025Affinity chromatography is the right answer
Neelam Sharma
September 23, 2025Affinity chromatography (step iv) stands as the most effective purification technique in this process due to its sharp increase in specific activity with minimal activity loss. Its molecular selectivity ensures superior enzyme enrichment compared to all other steps.
Aafreen Khan
September 23, 2025Affinity chromatography is best technique for enzyme purification