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Human Normal Immunoglobulin for Intramuscular Administration; IGIM

Category Passive immunizing agent.

          Immunoglobulin is a liquid or freeze-dried preparation containing immunoglobulins from normal human subjects, mainly immunoglobulin G (IgG). Other proteins may be present.

          Immunoglobulin is obtained from plasma complies with the requirements stated under Plasma for Fractionation, p. 193. No antibiotic is added to the plasma used.

Description The liquid Immunoglobulin is clear and pale-yellow to light-brown; during storage it may show formation of slight turbidity or a small amount of particulate matter.

The freeze-dried Immunoglobulin is a white or slightly yellow powder or solid, friable mass. It is hygroscopic.

Stability Liquid Immunoglobulin should be discarded if it has been frozen. The reconstituted solution of freeze-dried Immunoglobulin should be used immediately or as stated on the label.

Strengths available Liquid or reconstituted preparation, 150 to 180 mg of protein per ml.

Dose Intramuscular, preferably in the gluteal region, as directed by the physician.

Contra-indication

1. It is not for intravenous administration.

2. It is contra-indicated in patients with immunoglobulin A deficiencies since anaphylaxis may occur.

Warning

1. It may cause pain, tenderness and muscle stiffness at the site of injection.

2. It should be used with extreme caution in individuals with severe thrombocytopenia or any bleeding disorder.

3. Repeated injections of immunoglobulin, especially in allergic individuals, may result in sensitization which is usually manifested as fever, chills and sweating.

4. Risk-benefit should be considered if it is to be used in pregnant women.

Additional information The antibodies in immunoglobulin preparations may interfere with the immune response to certain live virus vaccines such as measles, mumps, rubella including MMR and varicella. These vaccines should be administered at least 2 to 3 weeks before or 3 months after treatment with immunoglobulin. However, there appears to be no interference between immunoglobulin and oral poliomyelitis vaccine (OPV), yellow fever vaccine, oral typhoid (strain Ty 21a) vaccine, or adsorbed diphtheria, pertussis (whole cell) and tetanus vaccine (DPT).

Expiration date When stored under the prescribed conditions, the expiration date is not later than 2 years from the date of manufacturing, or as indicated on the label.

Packaging and storage Liquid Immunoglobulin shall be kept in a sealed and colourless glass container, protected from light, and stored at a temperature of 2º to 8º; avoid freezing.

          Freeze-dried Immunoglobulin shall be kept in a tightly closed colourless glass container, protected from light, and stored at a temperature not exceeding 25º, unless otherwise specified by manufacturers.

Labelling Complies with the “General Information for Biological Products”, p. 177. In addition, the label on the container states (1) the volume and the total amount of the protein expressed in mg per ml or, for freezedried immunoglobulin, the total amount of protein in the container; (2) the route of administration; (3) where applicable, that the preparation is suitable for use in the prophylaxis of hepatitis A infection; (4) where applicable, the anti-hepatitis A virus activity in IU per ml. 

Before carrying out the identification and the tests (except those for constituted solution and water), immediately reconstitute the preparation to be examined as stated on the label.

 

Identification

          A. Carry out the precipitation tests on the preparation being examined, using an antiserum specific to human plasma proteins and a range of antisera specific to the plasma proteins of each species of domestic animal commonly used in the preparation of materials of biological origin. The preparation is shown to contain proteins of human origin and gives negative reactions with antisera specific to plasma proteins of other species.

          B. Examine by a suitable immunoelectrophoresis technique. Using antiserum to normal human serum, compare normal human serum and the preparation being examined, both diluted to contain 1 per cent w/v of protein. The main component of the preparation being examined corresponds to the IgG component of normal human serum. The solution may show the presence of small quantities of other plasma proteins.

pH 5.0 to 7.2, when diluted with saline TS to produce a solution containing 1 per cent w/v of protein (Appendix 4.11).

Total protein Not less than 10 per cent w/v and not more than 18 per cent w/v of protein and not less than 90 per cent and not more than 110 per cent of the labelled quantity of protein stated on the label. Dilute the preparation to be examined with saline TS to obtain a solution containing about 15 mg of protein in 2 ml. To 2.0 ml of this solution in a round-bottomed centrifuge tube add 2 ml of a 7.5 per cent w/v solution of sodium molybdate and 2 ml of a mixture of 1 volume of nitrogenfree sulfuric acid and 30 volumes of water. Shake, centrifuge for 5 minutes, decant the supernatant liquid, and allow the inverted tube to drain on filter paper. Using the residue thus obtained, carry out the “Determination of Nitrogen” (Method II, Appendix 6.7) and calculate the content of protein by multiplying by 6.25.

Protein composition Carry out the test as described in the “Cellulose Acetate Electrophoresis” (Method II, Appendix 3.7), but applying an electric field such that the albumin band of normal human serum applied in a control strip migrates at least 30 mm. Prepare the following solutions. For solution (A), dilute the preparation being examined with saline TS to produce a solution containing 5 per cent w/v of protein. For solution (B), dilute Human Immunoglobulin for Electrophoresis RS with saline TS to produce a solution containing 5 per cent w/v of protein. In the strips prepared from solution (A) not more than 10 per cent of the protein is contained in bands other than the principal band. The test is not valid unless the proportion of protein in the principal band in the strips prepared from solution (B) is within the limits stated in the leaflet supplied with Human Immunoglobulin for Electrophoresis RS.

Distribution of molecular size Carry out the test as described in the “High-pressure Liquid Chromatography” (Appendix 3.5).

          Mobile phase Dissolve and dilute 4.873 g of disodium hydrogenphosphate dihydrate, 1.741 g of sodium dihydrogenphosphate monohydrate, 11.688 g of sodium chloride and 50 mg of sodium azide with water to 1000.0 ml.

Standard preparation Dilute Human Immunoglobulin RS with saline TS to the same protein concentration as Test preparation.

          Test preparation Dilute the preparation being examined with saline TS to a concentration suitable for the chromatographic system used. A concentration in the range 0.4 to 1.2 per cent w/v of protein and the injection of 50 μg to 600 μg of protein are usuallysuitable.

          Chromatographic system The chromatographic procedure may be carried out using (a) a stainless steel column (60 cm × 7.5 mm) packed with hydrophilic silica gel (3 to 10 μm), (b) Mobile phase at a flow rate of about 0.5 ml per minute and (c) an ultraviolet photometer set at 280 nm.

          In the chromatogram obtained from Standard preparation, the principal peak corresponds to IgG monomer and there is a peak corresponding to dimer with a retention time relative to monomer of about 0.85. Identify the peaks in the chromatogram obtained from Test preparation by comparison with the chromatogram obtained from Standard preparation; any peak with retention time shorter than that of dimer corresponds to polymers and aggregates. The preparation to be examined complies with the test if, in the chromatogram obtained from Test preparation: for monomer and dimer, retention time relative to the corresponding peak in the chromatogram obtained from Standard preparation is 1±0.02; the sum of monomer and dimer represents not less than 85 per cent of the total area of the chromatogram and polymers and aggregates represent not more than 10 per cent of the total area of the chromatogram. This requirement does not apply to products where albumin has been added as a stabilizer; for products stabilized with albumin, a test for distribution of molecular size is carried out during manufacture before addition of the stabilizer.

Hemagglutinins, anti-A and anti-B Dilute the constituted solution with saline TS to produce a solution containing 3 per cent w/v of immunoglobulin. Carry out the test for hemagglutinins, anti-A and anti-B using a suitable indirect method such as that described below. The 1 in 64 dilutions do not show agglutination.

          Prepare in duplicates serial dilutions of the preparation being examined in saline TS. To each dilution of one series add an equal volume of a 5 per cent v/v suspension of group A1 red blood cells previously washed three times with saline TS. To each dilution of the other series add an equal volume of a 5 per cent v/v suspension of group B red blood cells previously washed three times with saline TS. Incubate the suspensions at 37º for 30 minutes and then wash the cells three times with saline TS. Leave the cells in contact with a polyvalent anti-human globulin reagent for 30 minutes. Without centrifuging, examine each suspension for agglutination under a microscope.

Anti-D antibodies If Immunoglobulin is intended for subcutaneous administration, it complies with the “Test for Anti-D Antibodies in Human Normal Immunoglobulin for Intravenous Administration” (Appendix 15.1.10).

Antibody to hepatitis B surface antigen Not less than 0.5 IU per g of immunoglobulin, determined by a suitable immunochemical method (Appendix 14.5).

Antibody to hepatitis A virus If intended for use in the prophylaxis of hepatitis A, it complies with the following additional requirement.

          Carry out the “Immunochemical Method” (Appendix 14.5). Determine the antibody content by comparison with a reference preparation calibrated in International Units, using an immunoassay of suitable sensitivity and specificity.

          The stated potency is not less than 100 IU per ml. The estimated potency is not less than the stated potency. The confidence limits (P = 0.95) are not less than 80 per cent and not more than 125 per cent of the estimated potency.

Sterility Complies with the “Sterility Test” (Method II, Appendix 10.1).

Pyrogens or Bacterial endotoxins Complies with the “Pyrogen Test” (Appendix 8.2) or, preferably and where justified and authorized, with a validated in vitro test such as the “Test for Bacterial Endotoxins” (Appendix 8.5).

          For the pyrogen test, use 1 ml of the preparation being examined per kg of the rabbit’s weight.

          Where the bacterial endotoxin test is used, it contains less than 5 Endotoxin Units per ml.

Other requirements Freeze-dried IGIM complies with the following additional requirements.

          Solubility test Add the volume of the liquid stated on the label. The preparation dissolves completely within 20 minutes at 20º to 25º.

          Water Not more than 3.0 per cent w/w (Karl Fischer Method, Appendix 4.12).