After usage the reagent should be immediately recapped

and replaced at 2–8°C storage.

ANTI-D (Rho) (IgG)

Monoclonal Blood Typing

Antibodies for Slide and Modified Tube Tests

(Courtesy: Tulip’s Eryclone Range)

Summary

Monoclonal antibodies are derived from hybridoma cell

lines, created by fusing mouse antibody producing Blymphocytes with mouse myeloma cells or are derived

from a human B cell line through EBV transformation. Each

hybridoma cell line produces homogeneous antibodies

of only one immunoglobulin class, which are identical in

their chemical structure and immunological activity.

Human red blood cells are classified as Rho(D) positive

or Rho(D) negative depending upon the presence or

absence of D(Rho) antigen on them. Approximately, 85%

of the Caucasian population is Rho(D) positive. The Du

phenotype is a variant of D(Rho) antigen and is recognized

by performing the antiglobulin test.

Reagent

Eryclone anti-D(Rho) (lgG) is a ready-to-use high-protein

reagent, prepared from supernatants of cell cultures with

Blood Banking (Immunohematology) 337

antibody producing B lymphocytes obtained through EBV

transformation. These antibodies of the immunoglobulin

class IgG are a mixture of several monoclonal antibodies

of the same specificity but having the capability of

recognizing different epitopes of the human red blood cell

antigen D (Rho).

Each batch of reagent undergoes rigorous quality

control at various stages of manufacture for its specificity,

avidity and performance.

Reagent Storage and Stability

1. Store the reagent at 2–8°C. Do not freeze.

2. The shelf life of reagent is as per the expiry date

mentioned on the reagent vial label.

Principle

Human red blood cells possessing the D(Rho) antigen

will agglutinate in the presence of antibody directed

towards the antigen. Agglutination of red blood cells with

Eryclone anti-D(Rho) (lgG) reagent is a positive test

result and indicates the presence of D(Rho) antigen. No

agglutination with anti-D(Rho) (lgG) reagent is a negative

test result and indicates the absence of D(Rho) antigen.

All negative test results should be further tested for Du

(Presence of weak/partial Ds) by performing the Du test

procedure as described later.

Note

1. In vitro diagnostic reagent for laboratory and

professional use only. Not for medicinal use.

2. Eryclone anti-D (Rho) (IgG) reagent is not from

human source, hence, contamination due to HBsAg

and HIV is practically excluded.

3. The reagent contains sodium azide 0.1% as preservative.

Avoid contact with skin and mucosa. On disposal flush

with large quantities of water.

4. Extreme turbidity may indicate microbial contamination or denaturation of protein due to thermal

damage. Such reagent should be discarded.

Sample Collection and Preparation

No special preparation of the patient is required prior

to sample collection by approved techniques. Samples

should be stored at 2–8°C if not tested immediately. Do not

use hemolyzed samples.

Anticoagulated blood using various anticoagulants

should be tested within the below mentioned time period

EDTA or heparin : 2 days

Sodium citrate or sodium oxalate : 14 days

ACD or CPD : 28 days.

Clotted whole blood should be tested within 14 days.

Additional Material Required for

Slide and Tube Tests

Glass slides (50 × 75 mm), test tubes (10 × 75 mm), Pasteur

pipettes, isotonic saline, centrifuge, timer, mixing sticks,