[2S] UNIT 6 Total Proteins

Question 1

first rank of importance

Answer 1

“Proteis”

Question 2

Consists of the elements carbon, oxygen, hydrogen, nitrogen, and sulfur (CHONS)

Answer 2

TOTAL PROTEIN

Question 2

Synthesized in the liver and secreted by the hepatocyte into the circulation except immunoglobulins

Answer 2

TOTAL PROTEIN

Question 2

can bear positive and negative charges (amphoteric)

Answer 2

Proteins

Question 3

pH range of protein

Answer 3

5.5 - 8.0

Question 4

T/F: Amino acids have no net charge (isoelectric point)

Answer 4

T

Question 5

pH > pI

Answer 5

Negative charge

Question 6

pH < pI

Answer 6

Positive charge

Question 7

Repair body tissues

Answer 7

Functions of Total Proteins

Question 8

For transport of metabolic substances

Answer 8

Functions of Total Proteins

Question 9

3. Hemostasis
4. Acid-base balance

Answer 9

Functions of Total Proteins

Question 10

5. Enzymes (biocatalysts)
6. Hormones
7. Structure (connective tissue)

Answer 10

Functions of Total Proteins

Question 11

Most frequently analyzed of all the proteins

Answer 11

PLASMA PROTEINS

Question 12

• Divided into two groups:
  
  • Albumins
  • Globulins

Answer 12

PLASMA PROTEINS

Question 12

TOTAL PROTEIN METHODS

Reference method. Assume average nitrogen content of
16%

Answer 12

Kjeldahl

Question 12

TOTAL PROTEIN METHODS

Measurement of refractive index (velocity of light in air and water) due to solutes in serum

Answer 12

Refractometry

Question 13

TOTAL PROTEIN METHODS

Formation of violet-colored chelate between Cu2+ ions
and peptide bonds (measured at 540 nm)

Answer 13

Biuret

Question 14

TOTAL PROTEIN METHODS

Protein binds to dye and causes a spectral shift in the absorbance maximum of dye

Answer 14

Dye Binding

Question 15

Acid precipitation (TCA or tungstic acid) of protein with measurement of
total nitrogen

Answer 15

Total Protein

Question 16

conversion of nitrogen to ammonia

Answer 16

Kjeldahlization

Question 16

2 Ammonia measurement

Answer 16

Nessler’s
Berthelot

Question 16

TOTAL PROTEIN METHODS

breaks the peptide bonds

Answer 16

BIURET: Cupric Ions

Question 17

TOTAL PROTEIN METHODS

stabilizes cupric ions

Answer 17

BIURET: Potassium iodide

Question 17

TOTAL PROTEIN METHODS

keeps copper in solution

Answer 17

BIURET: Tartrate Salt

Question 18

TOTAL PROTEIN METHODS 1. Bromphenol blue 2. Ponceau S 3. Amido black 10B 4. Lissamine green 5. Coomassie briliant blue

Answer 18

DYE BINDING

Question 19

* Synthesized in the liver * General transport protein

Answer 19

ALBUMIN

Question 20

It is the protein present in highest concentration in plasma

Answer 20

ALBUMIN

Question 21

Maintains osmotic pressure and is an indicator of nutritional status

Answer 21

ALBUMIN

Question 22

* Negative acute phase reactant * Lowest levels are seen in active Nephrotic syndrome

Answer 22

ALBUMIN

Question 23

ALBUMIN METHOD OF ANALYSIS Globulins are precipitated Albumin in supernatant is quantitated by biuret reaction

Answer 23

Salt Precipitation

Question 24

ALBUMIN METHOD OF ANALYSIS Nonspecific for Albumin

Answer 24

Methyl orange

Question 24

ALBUMIN METHOD OF ANALYSIS Sensitive Overestimates low albumin levels Most commonly used dye

Answer 24

BCG (Bromcresol green)

Question 24

ALBUMIN METHOD OF ANALYSIS Many Interferences (salicylates, bilirubin)

Answer 24

HABA

Question 25

ALBUMIN METHOD OF ANALYSIS Specific, Sensitive and Precise

Answer 25

BCP (Bromcresol purple)

Question 26

ALBUMIN METHOD OF ANALYSIS Proteins separated based on electric charge densities

Answer 26

Electrophoresis

Question 27

ALBUMIN METHOD OF ANALYSIS * Cellulose acetate/agarose gel (support media) * After separation, protein fractions are immersed in acid solution then stained by dyes (e.g. Coomassie blue)

Answer 27

Electrophoresis

Question 28

ALBUMIN METHOD OF ANALYSIS The medium is placed in scanning densitometer which compute the area under the absorbance

Answer 28

Electrophoresis

Question 29

ALBUMIN METHOD OF ANALYSIS Monoclonal increase

Answer 29

ELECTROPHORESIS: Monoclonal Gammopathy

Question 30

ALBUMIN METHOD OF ANALYSIS Uses higher voltage couple with a cooling system and more concentrated buffer

Answer 30

HIGH RESOLUTION PROTEIN ELECTROPHORESIS

Question 31

PRE-ANALYTICAL CONSIDERATIONS ❑ Non-fasting test ❑ Sample must be free from hemolysis (↑ increase) and lipemia

Answer 31

Total Proteins

Question 32

PRE-ANALYTICAL CONSIDERATIONS ❑ Concentration is higher in plasma than in serum * Presence of fibrinogen * Increased from 0.2 – 0.4 g/dL (2 – 4 g/L)

Answer 32

Total Proteins

Question 33

PROCEDURES Standard of TP

Answer 33

→ Protein: 8 g/dL or 80 g/L, → Sodium azide

Question 34

PROCEDURES TP Reagent Kit

Answer 34

→ Sodium Hydroxide → Potassium sodium tartrate → Copper sulfate → Potassium iodide

Question 35

PROCEDURES Incubation time for TP

Answer 35

10 minutes

Question 36

PROCEDURES Measure the absorbance of standard and the reagent blank within __ minutes

Answer 36

30

Question 36

PROCEDURES * Wavelength – 520 to 580 nm * Temperature – 37 degC * Optical path – 1 cm * Read against reagent blank

Answer 36

TP

Question 36

TOTAL PROTEIN: Increased / Decreased Dehydration

Answer 36

Increased

Question 37

TOTAL PROTEIN: Increased / Decreased * Chronic inflammation * Multiple myeloma

Answer 37

Increased

Question 38

TOTAL PROTEIN: Increased / Decreased * Nephrotic syndrome * Malabsorption

Answer 38

Decreased

Question 39

TOTAL PROTEIN: Increased / Decreased * Malnutrition * Agammaglobulinemia

Answer 39

Decreased

Question 39

TOTAL PROTEIN: Increased / Decreased * Overhydration * Hepatic insufficiency

Answer 39

Decreased

Question 39

ALBUMIN PROCEDURE Standard

Answer 39

Albumin 3.5 g/dL or 35 g/L

Question 39

REFERENCE VALUES (SERUM TOTAL PROTEIN)

Answer 39

6.5 – 8.3 g/dL (65-83 g/L)

Question 40

ALBUMIN PROCEDURE Albumin Reagent Kit

Answer 40

→ Succinate buffer (pH 4.2), → Bromcresol green → Surfactant → Sodium azide

Question 41

ALBUMIN PROCEDURE Incubation time

Answer 41

5 minutes

Question 42

ALBUMIN PROCEDURE Measure the absorbance of standard and the reagent blank within __ minutes

Answer 42

15

Question 43

PROCEDURE * Wavelength – 620 nm * Temperature – 37 degC * Optical path – 1 cm * Read against reagent blank

Answer 43

Albumin

Question 44

ALBUMIN: Increased / Decreased * Dehydration * Prolonged tourniquet application

Answer 44

Increased

Question 44

ALBUMIN: Increased / Decreased * Malnutrition * Liver disease

Answer 44

Decreased

Question 44

ALBUMIN: Increased / Decreased * Nephrotic syndrome * Chronic inflammation

Answer 44

Decreased

Question 45

REFERENCE VALUES (SERUM ALBUMIN)

Answer 45

3.5 – 5.5 g/dL (35-55 g/L)

Question 45

Computed using Total Protein and Albumin

Question 45

TP – Albumin =

Question 46

Normal value of Globulin

Answer 46

1.5 to 4.5 g/dL (15.0 to 45.0 g/L)

Question 47

GLOBULIN DETERMINATION A/G ratio

Answer 47

1.3 – 3.0

Question 48

GLOBULIN DETERMINATION Increased therapeutic drug in serum

Answer 48

Bisalbuminemia

Question 49

ODD ONE OUT: GLOBULIN DETERMINATION - Inverted A/G Ratio Cirrhosis Multiple Myeloma Waldenstroms Bisalbuminemia Macroglobulinemia

Answer 49

Bisalbuminemia

Question 49

TP: N/↓ A: ↓ G: ↑

Answer 49

Hepatic Damage & Infections

Question 50

HEPATIC DAMAGE TP: N/↓ A: ↓ G: ↑ β-γ bridging

Answer 50

Cirrhosis

Question 51

HEPATIC DAMAGE TP: N/↓ A: ↓ G: ↑ ↑ γ-globulins

Answer 51

Hepatitis

Question 52

INFECTIONS TP: N/↓ A: ↓ G: ↑ * ______ - α1 , α2 globulins * ______ - ↑ α1, α2 γ, globulins

Answer 52

Acute Chronic

Question 53

ODD ONE OUT: Electrophoresis TP: ↓ A: ↓ G: N Choices: Immunodeficiency Syndrome Inadequate diet Nephrotic syndrome ↑α2,β-globulins;↓γ-globulins

Answer 53

Immunodeficiency Syndrome

Question 54

TP: ↓ A: N G: ↓

Answer 54

Immunodeficiency syndrome

Question 55

TP: ↑ A: N G: ↑

Answer 55

Multiple myeloma Monoclonal & Polyclonal gammopathies

Question 55

TP: ↑ A: ↑ G: ↑

Answer 55

Dehydration