A Trial to Assess the Amount of Insulin Antibodies in Diabetic Patients by Surface Plasmon Resonance

Abstract

Objective To measure the amount and affinity of insulin antibodies, we performed a trial to establish a new method for quantitative and qualitative analysis of these antibodies by using surface plasmon resonance (BIAcore TM system). Methods Real-time detection of insulin antibody interaction and kinetic analysis were performed using the BIAcore TM system. Patients or Materials Eight diabetic patients with insulin antibodies and whose fasting total immunoreactive insulin levels were more than 100 U/ml were selected. The patients with and without recurrent hypoglycemia were classified into hypoglycemic episode-positive or hypoglycemic episode-negative groups, respectively. Seven diabetic patients without insulin antibodies were selected as controls.Results In the 8 patients, the concentration of insulin antibodies ranged from 2.91 to 16.3 g/ml and insulin antibodies were not detected in the control group. The apparent KD (dissociation constant) and kd (the dissociation rate constant) values of the patients were much larger than those seen for the anti-human insulin monoclonal antibody. The KD values were significantly higher in the hypoglycemic episode-positive group than in the hypoglycemic episode-negative group (p<0.05). No significant differences in the concentration, the ka (the association rate constant) and the kd values were noted between the groups.Conclusion The data suggests that insulin antibodies of the patients have an apparently lower affinity status in sera as compared with that for the anti-human insulin monoclonal antibody, and dissociate easily from the immune-complex in the sera, especially in cases where there is recurrent hypoglycemia in the patients. Therefore insulin antibody characteristics are one of the causative factors in hypoglycemic episodes. (Internal Medicine 44: 100-106, 2005)

Analytical and clinical challenges in a patient with concurrent type 1 diabetes, subcutaneous insulin resistance and insulin autoimmune syndrome

“…The characteristics of anti-insulin antibodies have been thoroughly studied in IAS. It is evident that high-affinity insulin antibodies frequently induce insulin resistance, whereas the presence of low-affinity antibodies induce hypoglycaemia due to the dissociation of the high amount of insulin from the insulin–antibody complex (12) (13). However, in diabetes, only a few studies have focused on the characteristics of these antibodies (6).…”

Immunological Responses to Exogenous Insulin

“…However, recent studies have reported potential progress toward identifying in vitro characteristics of IAs associated with clinical hypoglycemia syndromes in small numbers of patients (177,180,222,223,298). One of those studies has suggested that insulin-receiving individuals with high antibody levels and recurrent hypoglycemia have a higher dissociation constant for insulin measured by surface plasmon resonance than monoclonal antibodies to human insulin (298).…”

“…One of those studies has suggested that insulin-receiving individuals with high antibody levels and recurrent hypoglycemia have a higher dissociation constant for insulin measured by surface plasmon resonance than monoclonal antibodies to human insulin (298). It remains to be seen whether these recently proposed assays correlate better with symptoms than with standard IA measurements.…”

Circulating insulin antibodies: influence of continuous subcutaneous or intraperitoneal insulin infusion, and impact on glucose control

“…It must be presumed that antibodies first bind the insulin in circulation and then later dissociate from the insulin, allowing activation of cellular insulin receptors. While it was not clear that consistent affinity/binding capacity profiles can distinguish IAs in patients with and without hypoglycaemia syndromes [9], some progress has been reported recently [36,42]. For instance, insulinreceiving individuals with high IA levels and recurrent hypoglycaemia were shown to have a higher dissociation constant for insulin measured by surface plasmon resonance than monoclonal antibodies to human insulin [42].…”

“…While it was not clear that consistent affinity/binding capacity profiles can distinguish IAs in patients with and without hypoglycaemia syndromes [9], some progress has been reported recently [36,42]. For instance, insulinreceiving individuals with high IA levels and recurrent hypoglycaemia were shown to have a higher dissociation constant for insulin measured by surface plasmon resonance than monoclonal antibodies to human insulin [42]. In addition, many patients with type 1 diabetes have impaired glucose counter-regulation due to multiple defects in counter-regulatory hormone secretion, and the risk of severe hypoglycaemia was shown to be associated with increased IA binding, which prolonged the half-life of insulin [43,44].…”