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    Bel484 Major 04sem2

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    donotpanic
    This document contains the questions from a major exam in the course BE 484 Analytical Methods in Biotechnology. It includes 10 questions testing knowledge of topics like chromatography, electrophoresis, mass spectrometry, and radioisotope techniques. The questions require explanations of principles, calculations, and distinguishing between different analytical methods.

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    Bel484 Major 04sem2

    Uploaded by

    donotpanic
    133 views3 pages
    This document contains the questions from a major exam in the course BE 484 Analytical Methods in Biotechnology. It includes 10 questions testing knowledge of topics like chromatography, electrophoresis, mass spectrometry, and radioisotope techniques. The questions require explanations of principles, calculations, and distinguishing between different analytical methods.

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    This document contains the questions from a major exam in the course BE 484 Analytical Methods in Biotechnology. It includes 10 questions testing knowledge of topics like chromatography, electrophoresis, mass spectrometry, and radioisotope techniques. The questions require explanations of principles, calculations, and distinguishing between different analytical methods.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    133 views3 pages

    Bel484 Major 04sem2

    Uploaded by

    donotpanic
    This document contains the questions from a major exam in the course BE 484 Analytical Methods in Biotechnology. It includes 10 questions testing knowledge of topics like chromatography, electrophoresis, mass spectrometry, and radioisotope techniques. The questions require explanations of principles, calculations, and distinguishing between different analytical methods.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
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    BE 484 Analytical Methods in Biotechnology

    ,
    Major Exam (2ndsemester 2003-2004)

    AttemQLillJ ..9uestions

    Theory Max Marks: 40


    Time: Total 2 hours

    Q. I What is the basic principle of chromatographic method? Explain the difference


    between open and closed column chtomatography. Between HPLC and FPLC, which
    method has high resolving power and why? I + 1+ 2 = 4
    ':.t
    Q. 2 In a protein there are two solvent accessible tryptophan residues, one is on the
    ~4 surface and the other is in the interior surrounded by charged residues. How do you
    ;,
    selectively quench the tryptophan on the surface?Explain with principle the experiment. 2

    Q. 3 State and define various parameters that are used to evaluate the performance in
    chromu~0graphicseparations. 2
    ."
    Q. 4 Distinguish between g:-oupspecific ligand affinity chromatography and immobilized'
    metal affinity chromatography. Illustrate your answer with suitable examples. 3

    Q. 5 How does the polarity of the solvent affect the absorrtiuD maxima of the protein?
    How can the solvent rJerturbation method be' used to find the position of a particular
    an1inoacid in native protein? 3

    Q. b a. What is the principle on which zonal and isoUichl'phoresis electrophoretic


    systems are based? Gi"r. an application for each category. 3

    t... What is the function of the stacking and C separating gel 4\ystemsin DISC '.

    !f
    electrophoresis? .2
    <IiIr;"

    c. What are gradient gels and what might be these used for? 2

    Q. 7 a. Derive the expression relating velocity to mass-to-charge ratio in the MS


    method. An ion of mass m = 100 amu and charge z = 1, is accelerated with a potential of
    I kV. Calculate its velo<-ityl'l m/sec. The same ions encounter a magnet;..:field, the
    I

    geometry of which requires a trajectory with radius r = 10 cm for the ions to reachthe
    I exit slIt. What must be the magnetic field strength? 1+ 1+ 1= 3

    b. Why is it essential thC1f


    the ion-source be evacuated in the MS method?

    c. Distinguish between electron-impact ionization and chemical ionization. For


    which specific applications...would you use these
    . two different methods of ionization? 2
    -

    1
    't
    ~

    -j
    .~

    1
    y,

    !
    Laboratory Questions Max. Marks: 20 l
    !
    !
    I Q.1 a. While checking the punty nf a protein, which chromatographic system (FPLC or
    I HPLC) would you prefer and why? 2

    b. Proteins A and B interact in aqueous medium. How can you show with the HPLC
    experiment that they are binding to each other. Explain with thc drawings of the
    chromatogram. 2

    c. You have bC0ngiven two different concentrations of a pure protein for recording
    CD spectra. Would you get same results while caJculating its secondary structure? I
    2
    d.What is the need for having a pure protein in CD studies?

    Q.2 a. Why is methylation of a triglyceride carried out prior to GC analysis?

    b. State the reaction calalyzed by phospolipase A2. ...

    c.What is the principle of Folin-Lowry method for estimation of proteins in a


    fermentation (cell-free) broth?
    2
    d. Distinguish between transmittance and absorbance?
    2
    e. Name different parts of a typical double beam UY/visible spectrophotometer.

    Q.3 a. Proteins A and B having a molecular weight of 16,500 and 35,400 move 1.3 cm
    and 4.6 cm, respectively. whcn clecfrophore~ed through a gcL What is thc molecular
    weigh~of a protein C, which moves 2.8 cm in the saplp,gel? 2

    b. Wny is electrophoresis dOI1P;0 solutions of low sait concentration?

    c. A -,mallenzyme has a sedimentation coefficient:Jf 3.4 S. Whcn it bindS'to its


    substrate (a small organic molecule), its s value changes to 2.9 S. Explain this change. I

    d. If a boundary moves halfway down a centri(uge cell in 20 min at 20,000 rpm,


    how long w',:Jit take to reach the same position if the speed ;s 40.000 rpm ? 2

    - -
    --,---" "

    d. Show the ions being formed with methaneas a reagent gas. Would these be
    calledas the primaryor secondaryionsand why?Whatis theirfunction? 2+ 1+ 1 = 4

    Q.8 What is the difference between boundary and zonal sedimentation and under which
    !conditions(for what kind 0f data generation) would you use these two methods?
    ; 2+1= 3
    i
    'Q.9 a. The specific activity of a sample ofe2p] ATP is 5.3 Ci/mmol on Jan 23. What will
    2
    the specific activity be on Feb 18 of the same year~given the half-life of32p is 14.2days?

    b. In an experiment to measure DNA synthesis, would you use eH] thymidine or


    e2p] nucleotide and why? 1

    c. You have in your lab a Geiger counter with an efficiency of 22% for 14Cand a
    background of 6 cpm and a scintillation counter with an efficiency of 72% for 14Cand a
    background of 38 cpm. In an experiment with 14(:,you expect your sample to have very
    low activity-that is, from 75 to 100 cpm. Which counter should you chose andwhy? -3

    . -

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